Scholars@Duke
Aravind Asokan

Aravind Asokan

Professor in Surgery
Surgery, Surgical Sciences

Selected Publications

Progressive liver disease and dysregulated glycogen metabolism in murine GSD IX γ2 models human disease.

Journal Article Mol Genet Metab · December 2024 Hepatic glycogen storage disease type IX γ2 (GSD IX γ2) is a severe, liver-specific subtype of GSD IX. While all patients with hepatic GSD IX present with similar symptoms, over 95 % of patients with GSD IX γ2 progress to liver fibrosis and cirrhosis. Desp ... Full text Link to item Cite

Sub-genomic flaviviral RNA elements increase the stability and abundance of recombinant AAV vector transcripts.

Journal Article J Virol · August 20, 2024 UNLABELLED: Many viruses have evolved structured RNA elements that can influence transcript abundance and translational efficiency, and help evade host immune factors by hijacking cellular machinery during replication. Here, we evaluated the functional imp ... Full text Link to item Cite

Gene Delivery From Granular Scaffolds for Tunable Biologics Manufacturing.

Journal Article Small · August 2024 The understanding of the molecular basis for disease has generated a myriad of therapeutic biologics, including therapeutic proteins, antibodies, and viruses. However, the promise that biologics can resolve currently incurable diseases hinges in their manu ... Full text Link to item Cite

Evaluation of AAV Capsids and Delivery Approaches for Hereditary Hemorrhagic Telangiectasia Gene Therapy.

Journal Article Transl Stroke Res · July 8, 2024 Nosebleeds and intracranial hemorrhage from brain arteriovenous malformations (bAVMs) are among the most devastating symptoms of patients with hereditary hemorrhagic telangiectasis (HHT). All available managements have limitations. We showed that intraveno ... Full text Link to item Cite

Engineered IgM and IgG cleaving enzymes for mitigating antibody neutralization and complement activation in AAV gene transfer.

Journal Article Mol Ther · July 3, 2024 Systemic dosing of adeno-associated viral (AAV) vectors poses potential risk of adverse side effects including complement activation triggered by anti-capsid immunity. Due to the multifactorial nature of toxicities observed in this setting, a wide spectrum ... Full text Link to item Cite

Deep Learning Resolves Myovascular Dynamics in the Failing Human Heart.

Journal Article JACC Basic Transl Sci · May 2024 The adult mammalian heart harbors minute levels of cycling cardiomyocytes (CMs). Large numbers of images are needed to accurately quantify cycling events using microscopy-based methods. CardioCount is a new deep learning-based pipeline to rigorously score ... Full text Link to item Cite

Gene delivery followed by ex vivo lung perfusion using an adeno-associated viral vector in a rodent lung transplant model.

Journal Article J Thorac Cardiovasc Surg · May 2024 OBJECTIVE: Ex vivo lung perfusion has emerged as a platform for organ preservation, evaluation, and restoration. Gene delivery using a clinically relevant adeno-associated vector during ex vivo lung perfusion may be useful in optimizing donor allografts wh ... Full text Link to item Cite

Capsid-mediated control of adeno-associated viral transcription determines host range.

Journal Article Cell Rep · March 26, 2024 Adeno-associated virus (AAV) is a member of the genus Dependoparvovirus, which infects a wide range of vertebrate species. Here, we observe that, unlike most primate AAV isolates, avian AAV is transcriptionally silenced in human cells. By swapping the VP1  ... Full text Link to item Cite

Longitudinal intravital imaging of mouse placenta.

Journal Article Sci Adv · March 22, 2024 Studying placental functions is crucial for understanding pregnancy complications. However, imaging placenta is challenging due to its depth, volume, and motion distortions. In this study, we have developed an implantable placenta window in mice that enabl ... Full text Open Access Link to item Cite

Repurposing CRISPR-Cas13 systems for robust mRNA trans-splicing.

Journal Article Nat Commun · March 14, 2024 Type VI CRISPR enzymes have been developed as programmable RNA-guided Cas proteins for eukaryotic RNA editing. Notably, Cas13 has been utilized for site-targeted single base edits, demethylation, RNA cleavage or knockdown and alternative splicing. However, ... Full text Link to item Cite

A humanized mouse model for adeno-associated viral gene therapy.

Journal Article Nat Commun · March 4, 2024 Clinical translation of AAV-mediated gene therapy requires preclinical development across different experimental models, often confounded by variable transduction efficiency. Here, we describe a human liver chimeric transgene-free Il2rg-/-/Rag2-/-/Fah-/-/A ... Full text Link to item Cite

Engineering Synthetic circRNAs for Efficient CNS Expression.

Journal Article Methods Mol Biol · 2024 Circular RNAs (circRNAs) have recently emerged as a promising modality for gene and RNA-based therapies. They are more stable than their linear counterpart and can be designed for efficient expression in different cell and tissue types. In this chapter, we ... Full text Link to item Cite

Redirecting AAV vectors to extrahepatic tissues.

Journal Article Mol Ther · December 6, 2023 Recombinant adeno-associated viral (AAV) vectors are the current benchmark for systemic delivery of gene therapies to multiple organs in vivo. Despite clinical successes, safe and effective gene delivery to extrahepatic tissues has proven challenging due t ... Full text Link to item Cite

Structure-guided AAV capsid evolution strategies for enhanced CNS gene delivery.

Journal Article Nat Protoc · November 2023 Over the past 5 years, our laboratory has systematically developed a structure-guided library approach to evolve new adeno-associated virus (AAV) capsids with altered tissue tropism, higher transduction efficiency and the ability to evade pre-existing humo ... Full text Link to item Cite

Efficient Adeno-associated Virus-mediated Transgenesis in Alveolar Stem Cells and Associated Niches.

Journal Article Am J Respir Cell Mol Biol · September 2023 Targeted delivery of transgenes to tissue-resident stem cells and related niches offers avenues for interrogating pathways and editing endogenous alleles for therapeutic interventions. Here, we survey multiple adeno-associated virus (AAV) serotypes, admini ... Full text Link to item Cite

Spinal cord repair is modulated by the neurogenic factor Hb-egf under direction of a regeneration-associated enhancer.

Journal Article Nat Commun · August 11, 2023 Unlike adult mammals, zebrafish regenerate spinal cord tissue and recover locomotor ability after a paralyzing injury. Here, we find that ependymal cells in zebrafish spinal cords produce the neurogenic factor Hb-egfa upon transection injury. Animals with ... Full text Link to item Cite

AAV Gene Augmentation of Truncated Complement Factor H Differentially Rescues Ocular Complement Dysregulation in a Mouse Model.

Journal Article Invest Ophthalmol Vis Sci · July 3, 2023 PURPOSE: Complement dysregulation in the eye has been implicated in the pathogenesis of age-related macular degeneration (AMD), and genetic variants of complement factor H (CFH) are strongly associated with AMD risk. We therefore aimed to untangle the role ... Full text Link to item Cite

Adeno-associated virus mediates gene transduction after static cold storage treatment in rodent lung transplantation.

Conference J Thorac Cardiovasc Surg · July 2023 OBJECTIVE: Adeno-associated virus is a clinically used gene therapy vector but has not been studied in lung transplantation. We sought to determine the efficacy of adeno-associated virus delivery during static cold storage via the airway versus the pulmona ... Full text Link to item Cite

Interplay between Furin and Sialoglycans in Modulating Adeno-Associated Viral Cell Entry.

Journal Article J Virol · May 31, 2023 Adeno-associated viruses (AAVs) are small, helper-dependent, single-stranded DNA viruses that exploit a broad spectrum of host factors for cell entry. During the course of infection, several AAV serotypes have been shown to transit through the trans-Golgi ... Full text Link to item Cite

Rescue of glutaric aciduria type I in mice by liver-directed therapies.

Journal Article Sci Transl Med · April 19, 2023 Glutaric aciduria type I (GA-1) is an inborn error of metabolism with a severe neurological phenotype caused by the deficiency of glutaryl-coenzyme A dehydrogenase (GCDH), the last enzyme of lysine catabolism. Current literature suggests that toxic catabol ... Full text Link to item Cite

RESCUE OF GLUTARIC ACIDURIA TYPE I MICE BY LIVER DIRECTED THERAPIES

Conference Molecular Genetics and Metabolism · March 2023 Full text Cite

An evolved AAV variant enables efficient genetic engineering of murine T cells.

Journal Article Cell · January 19, 2023 Precise targeting of large transgenes to T cells using homology-directed repair has been transformative for adoptive cell therapies and T cell biology. Delivery of DNA templates via adeno-associated virus (AAV) has greatly improved knockin efficiencies, bu ... Full text Link to item Cite

An enhancer-based gene-therapy strategy for spatiotemporal control of cargoes during tissue repair.

Journal Article Cell Stem Cell · January 5, 2023 The efficacy and safety of gene-therapy strategies for indications like tissue damage hinge on precision; yet, current methods afford little spatial or temporal control of payload delivery. Here, we find that tissue-regeneration enhancer elements (TREEs) i ... Full text Link to item Cite

AAV9 PD-L1 Mediated Immunodulation of Donor Graft in Rat Lung Allotransplantation

Conference JOURNAL OF HEART AND LUNG TRANSPLANTATION · 2023 Cite

CRISPR Assisted Trans-Splicing of RNA Fragments

Conference MOLECULAR THERAPY · 2023 Cite

Evaluating AAV vectors for HHT gene therapy

Conference ANGIOGENESIS · 2023 Cite

Multi-apical polarity of alveolar stem cells and their dynamics during lung development and regeneration.

Journal Article iScience · October 21, 2022 Epithelial cells of diverse tissues are characterized by the presence of a single apical domain. In the lung, electron microscopy studies have suggested that alveolar type-2 epithelial cells (AT2s) en face multiple alveolar sacs. However, apical and basola ... Full text Open Access Link to item Cite

Cross-species evolution of a highly potent AAV variant for therapeutic gene transfer and genome editing.

Journal Article Nat Commun · October 10, 2022 Recombinant adeno-associated viral (AAV) vectors are a promising gene delivery platform, but ongoing clinical trials continue to highlight a relatively narrow therapeutic window. Effective clinical translation is confounded, at least in part, by difference ... Full text Link to item Cite

Epigenetic Silencing of Recombinant Adeno-associated Virus Genomes by NP220 and the HUSH Complex.

Journal Article J Virol · February 23, 2022 The single-stranded DNA genome of adeno-associated viruses (AAV) undergoes second-strand synthesis and transcription in the host cell nucleus. While wild-type AAV genomes are naturally silenced upon integration into the host genome, recombinant AAV (rAAV) ... Full text Link to item Cite

Structurally Mapping Antigenic Epitopes of Adeno-associated Virus 9: Development of Antibody Escape Variants.

Journal Article J Virol · February 9, 2022 Adeno-associated viruses (AAV) serve as vectors for therapeutic gene delivery. AAV9 vectors have been FDA approved, as Zolgensma, for the treatment of spinal muscular atrophy and are being evaluated in clinical trials for the treatment of neurotropic and m ... Full text Link to item Cite

Targeted Delivery for Cardiac Regeneration: Comparison of Intra-coronary Infusion and Intra-myocardial Injection in Porcine Hearts.

Journal Article Front Cardiovasc Med · 2022 BACKGROUND: The optimal delivery route to enhance effectiveness of regenerative therapeutics to the human heart is poorly understood. Direct intra-myocardial (IM) injection is the gold standard, however, it is relatively invasive. We thus compared targeted ... Full text Link to item Cite

Gene Therapy: Will the Promise of Optimizing Lung Allografts Become Reality?

Journal Article Front Immunol · 2022 Lung transplantation is the definitive therapy for patients living with end-stage lung disease. Despite significant progress made in the field, graft survival remains the lowest of all solid organ transplants. Additionally, the lung has among the lowest of ... Full text Link to item Cite

Full-length dystrophin restoration via targeted exon integration by AAV-CRISPR in a humanized mouse model of Duchenne muscular dystrophy.

Journal Article Mol Ther · November 3, 2021 Targeted gene-editing strategies have emerged as promising therapeutic approaches for the permanent treatment of inherited genetic diseases. However, precise gene correction and insertion approaches using homology-directed repair are still limited by low e ... Full text Link to item Cite

The membrane associated accessory protein is an adeno-associated viral egress factor.

Journal Article Nat Commun · October 29, 2021 Adeno-associated viruses (AAV) rely on helper viruses to transition from latency to lytic infection. Some AAV serotypes are secreted in a pre-lytic manner as free or extracellular vesicle (EV)-associated particles, although mechanisms underlying such are u ... Full text Link to item Cite

Receptor Switching in Newly Evolved Adeno-associated Viruses.

Journal Article J Virol · September 9, 2021 Adeno-associated viruses utilize different glycans and the AAV receptor (AAVR) for cellular attachment and entry. Directed evolution has yielded new AAV variants; however, structure-function correlates underlying their improved transduction are generally o ... Full text Link to item Cite

Transgenic mice for in vivo epigenome editing with CRISPR-based systems.

Journal Article Nat Methods · August 2021 CRISPR-Cas9 technologies have dramatically increased the ease of targeting DNA sequences in the genomes of living systems. The fusion of chromatin-modifying domains to nuclease-deactivated Cas9 (dCas9) has enabled targeted epigenome editing in both culture ... Full text Link to item Cite

Characterization of liver GSD IX γ2 pathophysiology in a novel Phkg2-/- mouse model.

Journal Article Mol Genet Metab · July 2021 INTRODUCTION: Liver Glycogen Storage Disease IX is a rare metabolic disorder of glycogen metabolism caused by deficiency of the phosphorylase kinase enzyme (PhK). Variants in the PHKG2 gene, encoding the liver-specific catalytic γ2 subunit of PhK, are asso ... Full text Open Access Link to item Cite

AAV-CNS matters turn from gray to white.

Journal Article Mol Ther · May 5, 2021 Full text Link to item Cite

Adeno-Associated Virus-Mediated Gene Therapy in the Mashlool, Atp1a3Mashl/+, Mouse Model of Alternating Hemiplegia of Childhood.

Journal Article Hum Gene Ther · April 2021 Alternating Hemiplegia of Childhood (AHC) is a devastating autosomal dominant disorder caused by ATP1A3 mutations, resulting in severe hemiplegia and dystonia spells, ataxia, debilitating disabilities, and premature death. Here, we determine the effects of ... Full text Link to item Cite

The NIH Somatic Cell Genome Editing program.

Journal Article Nature · April 2021 The move from reading to writing the human genome offers new opportunities to improve human health. The United States National Institutes of Health (NIH) Somatic Cell Genome Editing (SCGE) Consortium aims to accelerate the development of safer and more-eff ... Full text Link to item Cite

A Beautiful Mind and the Heart of an Explorer

Journal Article Human Gene Therapy · April 1, 2021 Full text Cite

Engineering highly efficient backsplicing and translation of synthetic circRNAs.

Journal Article Mol Ther Nucleic Acids · March 5, 2021 Circular RNAs (circRNAs) are highly stable RNA molecules that are attractive templates for expression of therapeutic proteins and non-coding RNAs. In eukaryotes, circRNAs are primarily generated by the spliceosome through backsplicing. Here, we interrogate ... Full text Link to item Cite

Safe and Effective In Vivo Targeting and Gene Editing in Hematopoietic Stem Cells: Strategies for Accelerating Development.

Journal Article Hum Gene Ther · January 2021 On May 11, 2020, the National Institutes of Health (NIH) and the Bill & Melinda Gates Foundation (Gates Foundation) held an exploratory expert scientific roundtable to inform an NIH-Gates Foundation collaboration on the development of scalable, sustainable ... Full text Link to item Cite

In Vivo Gene Editing of Muscle Stem Cells with Adeno-Associated Viral Vectors in a Mouse Model of Duchenne Muscular Dystrophy.

Journal Article Mol Ther Methods Clin Dev · December 11, 2020 Delivery of therapeutic transgenes with adeno-associated viral (AAV) vectors for treatment of myopathies has yielded encouraging results in animal models and early clinical studies. Although certain AAV serotypes efficiently target muscle fibers, transduct ... Full text Link to item Cite

Optimizing delivery for efficient cardiac reprogramming.

Journal Article Biochem Biophys Res Commun · November 26, 2020 Following heart injury, cardiomyocytes, are lost and are not regenerated. In their place, fibroblasts invade the dead tissue where they generate a scar, which reduces cardiac function. We and others have demonstrated that combinations of specific miRNAs (m ... Full text Link to item Cite

The Golgi Calcium ATPase Pump Plays an Essential Role in Adeno-associated Virus Trafficking and Transduction.

Journal Article J Virol · October 14, 2020 Adeno-associated viruses (AAVs) are dependoparvoviruses that have proven useful for therapeutic gene transfer; however, our understanding of host factors that influence AAV trafficking and transduction is still evolving. Here, we investigated the role of c ... Full text Link to item Cite

Rescuing AAV gene transfer from neutralizing antibodies with an IgG-degrading enzyme.

Journal Article JCI Insight · September 17, 2020 Preexisting humoral immunity to recombinant adeno-associated virus (AAV) vectors restricts the treatable patient population and efficacy of human gene therapies. Approaches to clear neutralizing antibodies (NAbs), such as plasmapheresis and immunosuppressi ... Full text Link to item Cite

Coevolution of Adeno-associated Virus Capsid Antigenicity and Tropism through a Structure-Guided Approach.

Journal Article J Virol · September 15, 2020 Adeno-associated viruses (AAV) are composed of nonenveloped, icosahedral protein shells that can be adapted to package and deliver recombinant therapeutic DNA. Approaches to engineer recombinant capsids for gene therapy applications have focused on rationa ... Full text Link to item Cite

Gangliosides are essential endosomal receptors for quasi-enveloped and naked hepatitis A virus.

Journal Article Nat Microbiol · September 2020 The Picornaviridae are a diverse family of positive-strand RNA viruses that includes numerous human and veterinary pathogens1. Among these, hepatitis A virus (HAV), a common cause of acute hepatitis in humans, is unique in that it is hepatotropic and is re ... Full text Link to item Cite

Defining Transcription Regulatory Elements in the Human Frataxin Gene: Implications for Gene Therapy.

Journal Article Hum Gene Ther · August 2020 Friedreich's ataxia (FRDA) is the most common inherited form of ataxia in humans. It is caused by severe downregulation of frataxin (FXN) expression instigated by hyperexpansion of the GAA repeats located in intron 1 of the FXN gene. Despite numerous studi ... Full text Link to item Cite

Intravital imaging of mouse embryos.

Journal Article Science · April 10, 2020 Embryonic development is a complex process that is unamenable to direct observation. In this study, we implanted a window to the mouse uterus to visualize the developing embryo from embryonic day 9.5 to birth. This removable intravital window allowed manip ... Full text Open Access Link to item Cite

A CRISPR Screen Identifies the Cell Polarity Determinant Crumbs 3 as an Adeno-associated Virus Restriction Factor in Hepatocytes.

Journal Article J Virol · November 1, 2019 Adeno-associated viruses (AAV) are helper-dependent parvoviruses that have been developed into promising gene therapy vectors. Many studies, including a recent unbiased genomic screen, have identified host factors essential for AAV cell entry, but no genom ... Full text Link to item Cite

Vaccine-Mediated Inhibition of the Transporter Associated with Antigen Processing Is Insufficient To Induce Major Histocompatibility Complex E-Restricted CD8+ T Cells in Nonhuman Primates.

Journal Article J Virol · October 1, 2019 Major histocompatibility complex E (MHC-E) is a highly conserved nonclassical MHC-Ib molecule that tightly binds peptides derived from leader sequences of classical MHC-Ia molecules for presentation to natural killer cells. However, MHC-E also binds divers ... Full text Link to item Cite

Ring finger protein 121 is a potent regulator of adeno-associated viral genome transcription.

Journal Article PLoS Pathog · August 2019 Adeno-associated viruses (AAV) are Dependoparvoviruses that have shown promise as recombinant vectors for gene therapy. While infectious pathways of AAV are well studied, gaps remain in our understanding of host factors affecting vector genome expression. ... Full text Link to item Cite

Modulation of Sialic Acid Dependence Influences the Central Nervous System Transduction Profile of Adeno-associated Viruses.

Journal Article J Virol · June 1, 2019 Central nervous system (CNS) transduction by systemically administered recombinant adeno-associated viral (AAV) vectors requires crossing the blood-brain barrier (BBB). We recently mapped a structural footprint on the AAVrh.10 capsid, which, when grafted o ... Full text Link to item Cite

Long-term evaluation of AAV-CRISPR genome editing for Duchenne muscular dystrophy.

Journal Article Nat Med · March 2019 Duchenne muscular dystrophy (DMD) is a monogenic disorder and a candidate for therapeutic genome editing. There have been several recent reports of genome editing in preclinical models of Duchenne muscular dystrophy1-6, however, the long-term persistence a ... Full text Link to item Cite

Aquaporin-4-dependent glymphatic solute transport in the rodent brain.

Journal Article Elife · December 18, 2018 The glymphatic system is a brain-wide clearance pathway; its impairment contributes to the accumulation of amyloid-β. Influx of cerebrospinal fluid (CSF) depends upon the expression and perivascular localization of the astroglial water channel aquaporin-4 ... Full text Link to item Cite

Tissue-Dependent Expression and Translation of Circular RNAs with Recombinant AAV Vectors In Vivo.

Journal Article Mol Ther Nucleic Acids · December 7, 2018 Circular RNAs (circRNAs) are long-lived, covalently closed RNAs that are abundantly expressed and evolutionarily conserved across eukaryotes. Possible functions ranging from microRNA (miRNA) and RNA binding protein sponges to regulators of transcription an ... Full text Link to item Cite

Physical positioning markedly enhances brain transduction after intrathecal AAV9 infusion.

Journal Article Sci Adv · November 2018 Several neurological disorders may benefit from gene therapy. However, even when using the lead vector candidate for intrathecal administration, adeno-associated virus serotype 9 (AAV9), the strength and distribution of gene transfer to the brain are incon ... Full text Link to item Cite

Mapping and Engineering Functional Domains of the Assembly-Activating Protein of Adeno-associated Viruses.

Journal Article J Virol · July 15, 2018 Adeno-associated viruses (AAVs) encode a unique assembly-activating protein (AAP) within their genomes that is essential for capsid assembly. Studies to date have focused on establishing the role of AAP as a chaperone that mediates the stability, nucleolar ... Full text Link to item Cite

An Adeno-Associated Virus-Based Toolkit for Preferential Targeting and Manipulating Quiescent Neural Stem Cells in the Adult Hippocampus.

Journal Article Stem Cell Reports · March 13, 2018 Quiescent neural stem cells (qNSCs) with radial morphology are the only proven source of new neurons in the adult mammalian brain. Our understanding of the roles of newly generated neurons depends on the ability to target and manipulate adult qNSCs. Althou ... Full text Link to item Cite

Mapping the Structural Determinants Required for AAVrh.10 Transport across the Blood-Brain Barrier.

Journal Article Mol Ther · February 7, 2018 Effective gene delivery to the CNS by intravenously administered adeno-associated virus (AAV) vectors requires crossing the blood-brain barrier (BBB). To achieve therapeutic CNS transgene expression, high systemic vector doses are often required, which pos ... Full text Link to item Cite

Hepatocytic expression of human sodium-taurocholate cotransporting polypeptide enables hepatitis B virus infection of macaques.

Journal Article Nat Commun · December 15, 2017 Hepatitis B virus (HBV) is a major global health concern, and the development of curative therapeutics is urgently needed. Such efforts are impeded by the lack of a physiologically relevant, pre-clinical animal model of HBV infection. Here, we report that ... Full text Link to item Cite

Primary Cilia Signaling Shapes the Development of Interneuronal Connectivity.

Journal Article Dev Cell · August 7, 2017 Appropriate growth and synaptic integration of GABAergic inhibitory interneurons are essential for functional neural circuits in the brain. Here, we demonstrate that disruption of primary cilia function following the selective loss of ciliary GTPase Arl13b ... Full text Link to item Cite

Structure-guided evolution of antigenically distinct adeno-associated virus variants for immune evasion.

Journal Article Proc Natl Acad Sci U S A · June 13, 2017 Preexisting neutralizing antibodies (NAbs) against adeno-associated viruses (AAVs) pose a major, unresolved challenge that restricts patient enrollment in gene therapy clinical trials using recombinant AAV vectors. Structural studies suggest that despite a ... Full text Link to item Cite

Systemic and Persistent Muscle Gene Expression in Rhesus Monkeys with a Liver De-Targeted Adeno-Associated Virus Vector.

Journal Article Hum Gene Ther · May 2017 The liver is a major off-target organ in gene therapy approaches for cardiac and musculoskeletal disorders. Intravenous administration of most of the naturally occurring adeno-associated virus (AAV) strains invariably results in vector genome sequestration ... Full text Link to item Cite

Inducing circular RNA formation using the CRISPR endoribonuclease Csy4.

Journal Article RNA · May 2017 Circular RNAs (circRNAs) are highly stable, covalently closed RNAs that are regulated in a spatiotemporal manner and whose functions are largely unknown. These molecules have the potential to be incorporated into engineered systems with broad technological ... Full text Link to item Cite

Probing the Link among Genomic Cargo, Contact Mechanics, and Nanoindentation in Recombinant Adeno-Associated Virus 2.

Journal Article J Phys Chem B · March 2, 2017 Recombinant adeno-associated virus (AAV) is a promising gene therapy vector. To make progress in this direction, the relationship between the characteristics of the genomic cargo and the capsid stability must be understood in detail. The goal of this study ... Full text Link to item Cite

Cellular transduction mechanisms of adeno-associated viral vectors.

Journal Article Curr Opin Virol · December 2016 Recombinant adeno-associated viral vectors (rAAV) are regarded as promising vehicles for therapeutic gene delivery. Continued development and new strategies are essential to improve the potency of AAV vectors and reduce the effective dose needed for clinic ... Full text Link to item Cite

Generation and characterization of anti-Adeno-associated virus serotype 8 (AAV8) and anti-AAV9 monoclonal antibodies.

Journal Article J Virol Methods · October 2016 Adeno-associated viruses (AAVs) are promising viral vectors for therapeutic gene delivery, and the approval of an AAV1 vector for the treatment of lipoprotein lipase deficiency has heralded a new and exciting era for this system. However, preclinical and c ... Full text Link to item Cite

Glymphatic fluid transport controls paravascular clearance of AAV vectors from the brain.

Journal Article JCI Insight · September 8, 2016 Adeno-associated viruses (AAV) are currently being evaluated in clinical trials for gene therapy of CNS disorders. However, host factors that influence the spread, clearance, and transduction efficiency of AAV vectors in the brain are not well understood. ... Full text Link to item Cite

CNS-restricted Transduction and CRISPR/Cas9-mediated Gene Deletion with an Engineered AAV Vector.

Journal Article Mol Ther Nucleic Acids · July 19, 2016 Gene therapy using recombinant adeno-associated viral (AAV) vectors is emerging as a promising approach to treat central nervous system disorders such as Spinal muscular atrophy, Batten, Parkinson and Alzheimer disease amongst others. A critical remaining ... Full text Link to item Cite

Resolving Adeno-Associated Viral Particle Diversity With Charge Detection Mass Spectrometry.

Journal Article Anal Chem · July 5, 2016 Recombinant adeno-associated viruses (AAVs) are promising vectors for human gene therapy. However, current methods for evaluating AAV particle populations and vector purity are inefficient and low resolution. Here, we show that charge detection mass spectr ... Full text Link to item Cite

Engineering AAV receptor footprints for gene therapy.

Journal Article Curr Opin Virol · June 2016 Adeno-associated viruses (AAV) are currently at the forefront of human gene therapy clinical trials as recombinant vectors. Significant progress has been made in elucidating the structure, biology and tropisms of different naturally occurring AAV isolates ... Full text Link to item Cite

Characterization of the Adeno-Associated Virus 1 and 6 Sialic Acid Binding Site.

Journal Article J Virol · June 1, 2016 UNLABELLED: The adeno-associated viruses (AAVs), which are being developed as gene delivery vectors, display differential cell surface glycan binding and subsequent tissue tropisms. For AAV serotype 1 (AAV1), the first viral vector approved as a gene thera ... Full text Link to item Cite

rAAV-compatible MiniPromoters for restricted expression in the brain and eye.

Journal Article Mol Brain · May 10, 2016 BACKGROUND: Small promoters that recapitulate endogenous gene expression patterns are important for basic, preclinical, and now clinical research. Recently, there has been a promising revival of gene therapy for diseases with unmet therapeutic needs. To da ... Full text Link to item Cite

AAV Gene Therapy for MPS1-associated Corneal Blindness.

Journal Article Sci Rep · February 22, 2016 Although cord blood transplantation has significantly extended the lifespan of mucopolysaccharidosis type 1 (MPS1) patients, over 95% manifest cornea clouding with about 50% progressing to blindness. As corneal transplants are met with high rejection rates ... Full text Open Access Link to item Cite

Development of Patient-specific AAV Vectors After Neutralizing Antibody Selection for Enhanced Muscle Gene Transfer.

Journal Article Mol Ther · February 2016 A major hindrance in gene therapy trials with adeno-associated virus (AAV) vectors is the presence of neutralizing antibodies (NAbs) that inhibit AAV transduction. In this study, we used directed evolution techniques in vitro and in mouse muscle to select ... Full text Link to item Cite

In vivo genome editing improves muscle function in a mouse model of Duchenne muscular dystrophy.

Journal Article Science · January 22, 2016 Duchenne muscular dystrophy (DMD) is a devastating disease affecting about 1 out of 5000 male births and caused by mutations in the dystrophin gene. Genome editing has the potential to restore expression of a modified dystrophin gene from the native locus ... Full text Link to item Cite

Chemical Modulation of Endocytic Sorting Augments Adeno-associated Viral Transduction.

Journal Article J Biol Chem · January 8, 2016 Intracellular trafficking of viruses can be influenced by a variety of inter-connected cellular sorting and degradation pathways involving endo-lysosomal vesicles, the ubiquitin-proteasome system, and autophagy-based or endoplasmic reticulum-associated mac ... Full text Link to item Cite

Gene therapy of CNS disorders using recombinant AAV vectors

Chapter · January 1, 2016 Corrective intervention for CNS disorders typically requires replenishment of depleted biomolecules (e.g., catabolic enzymes), protection of neurons and glia from premature death, or utilization of CNS cells as bio-factories for production of neurotransmit ... Full text Cite

Controlling mRNA stability and translation with the CRISPR endoribonuclease Csy4.

Journal Article RNA · November 2015 The bacterial CRISPR endoribonuclease Csy4 has recently been described as a potential RNA processing tool. Csy4 recognizes substrate RNA through a specific 28-nt hairpin sequence and cleaves at the 3' end of the stem. To further explore applicability in ma ... Full text Link to item Cite

An siRNA Screen Identifies the U2 snRNP Spliceosome as a Host Restriction Factor for Recombinant Adeno-associated Viruses.

Journal Article PLoS Pathog · August 2015 Adeno-associated viruses (AAV) have evolved to exploit the dynamic reorganization of host cell machinery during co-infection by adenoviruses and other helper viruses. In the absence of helper viruses, host factors such as the proteasome and DNA damage resp ... Full text Link to item Cite

Strategies to circumvent humoral immunity to adeno-associated viral vectors.

Journal Article Expert Opin Biol Ther · June 2015 INTRODUCTION: Recent success in gene therapy of certain monogenic diseases in the clinic has infused enthusiasm into the continued development of recombinant adeno-associated viral (AAV) vectors as next-generation biologics. However, progress in clinical t ... Full text Link to item Cite

Optical Control of CRISPR/Cas9 Gene Editing.

Journal Article J Am Chem Soc · May 6, 2015 The CRISPR/Cas9 system has emerged as an important tool in biomedical research for a wide range of applications, with significant potential for genome engineering and gene therapy. In order to achieve conditional control of the CRISPR/Cas9 system, a geneti ... Full text Link to item Cite

Unique glycan signatures regulate adeno-associated virus tropism in the developing brain.

Journal Article J Virol · April 2015 UNLABELLED: Adeno-associated viruses (AAV) are thought to spread through the central nervous system (CNS) by exploiting cerebrospinal fluid (CSF) flux and hijacking axonal transport pathways. The role of host receptors that mediate these processes is not w ... Full text Link to item Cite

Functional analysis of the putative integrin recognition motif on adeno-associated virus 9.

Journal Article J Biol Chem · January 16, 2015 Adeno-associated viruses (AAVs) display a highly conserved NGR motif on the capsid surface. Earlier studies have established this tripeptide motif as being essential for integrin-mediated uptake of recombinant AAV serotype 2 (AAV2) in cultured cells. Howev ... Full text Link to item Cite

Cardiac I-1c overexpression with reengineered AAV improves cardiac function in swine ischemic heart failure.

Journal Article Mol Ther · December 2014 Cardiac gene therapy has emerged as a promising option to treat advanced heart failure (HF). Advances in molecular biology and gene targeting approaches are offering further novel options for genetic manipulation of the cardiovascular system. The aim of th ... Full text Link to item Cite

Preclinical toxicity evaluation of AAV for pain: evidence from human AAV studies and from the pharmacology of analgesic drugs.

Journal Article Mol Pain · September 2, 2014 Gene therapy with adeno-associated virus (AAV) has advanced in the last few years from promising results in animal models to >100 clinical trials (reported or under way). While vector availability was a substantial hurdle a decade ago, innovative new produ ... Full text Link to item Cite

Gene therapy: charting a future course--summary of a National Institutes of Health Workshop, April 12, 2013.

Journal Article Hum Gene Ther · June 2014 Recently, the gene therapy field has begun to experience clinical successes in a number of different diseases using various approaches and vectors. The workshop Gene Therapy: Charting a Future Course, sponsored by the National Institutes of Health (NIH) Of ... Full text Link to item Cite

Biology of adeno-associated viral vectors in the central nervous system.

Journal Article Front Mol Neurosci · 2014 Gene therapy is a promising approach for treating a spectrum of neurological and neurodegenerative disorders by delivering corrective genes to the central nervous system (CNS). In particular, adeno-associated viruses (AAVs) have emerged as promising tools ... Full text Link to item Cite

Multiple roles for sialylated glycans in determining the cardiopulmonary tropism of adeno-associated virus 4.

Journal Article J Virol · December 2013 Adeno-associated virus 4 (AAV4) is one of the most divergent serotypes among known AAV isolates. Mucins or O-linked sialoglycans have been identified as the primary attachment receptors for AAV4 in vitro. However, little is known about the role(s) played b ... Full text Link to item Cite

An emerging adeno-associated viral vector pipeline for cardiac gene therapy.

Journal Article Hum Gene Ther · November 2013 The naturally occurring adeno-associated virus (AAV) isolates display diverse tissue tropisms in different hosts. Robust cardiac transduction in particular has been reported for certain AAV strains. Successful applications of these AAV strains in preclinic ... Full text Link to item Cite

Engraftment of a galactose receptor footprint onto adeno-associated viral capsids improves transduction efficiency.

Journal Article J Biol Chem · October 4, 2013 New viral strains can be evolved to recognize different host glycans through mutagenesis and experimental adaptation. However, such mutants generally harbor amino acid changes that affect viral binding to a single class of carbohydrate receptors. We descri ... Full text Link to item Cite

Biophysical and ultrastructural characterization of adeno-associated virus capsid uncoating and genome release.

Journal Article J Virol · March 2013 We describe biophysical and ultrastructural differences in genome release from adeno-associated virus (AAV) capsids packaging wild-type DNA, recombinant single-stranded DNA (ssDNA), or dimeric, self-complementary DNA (scDNA) genomes. Atomic force microscop ... Full text Link to item Cite

Glycan binding avidity determines the systemic fate of adeno-associated virus type 9.

Journal Article J Virol · October 2012 Glycans are key determinants of host range and transmissibility in several pathogens. In the case of adeno-associated viruses (AAV), different carbohydrates serve as cellular receptors in vitro; however, their contributions in vivo are less clear. A partic ... Full text Link to item Cite

Single amino acid modification of adeno-associated virus capsid changes transduction and humoral immune profiles.

Journal Article J Virol · August 2012 Adeno-associated virus (AAV) vectors have the potential to promote long-term gene expression. Unfortunately, humoral immunity restricts patient treatment and in addition provides an obstacle to the potential option of vector readministration. In this study ... Full text Link to item Cite

Tyrosine cross-linking reveals interfacial dynamics in adeno-associated viral capsids during infection.

Journal Article ACS Chem Biol · June 15, 2012 Viral capsid dynamics are often observed during infectious events such as cell surface attachment, entry and genome release. Structural analysis of adeno-associated virus (AAV), a helper-dependent parvovirus, revealed a cluster of surface-exposed tyrosine ... Full text Link to item Cite

Human galectin 3 binding protein interacts with recombinant adeno-associated virus type 6.

Journal Article J Virol · June 2012 Recombinant adeno-associated viruses (rAAVs) hold enormous potential for human gene therapy. Despite the well-established safety and efficacy of rAAVs for in vivo gene transfer, there is still little information concerning the fate of vectors in blood foll ... Full text Link to item Cite

The AAV vector toolkit: poised at the clinical crossroads.

Journal Article Mol Ther · April 2012 The discovery of naturally occurring adeno-associated virus (AAV) isolates in different animal species and the generation of engineered AAV strains using molecular genetics tools have yielded a versatile AAV vector toolkit. Promising results in preclinical ... Full text Link to item Cite

Neutralizing antibodies against adeno-associated virus examined prospectively in pediatric patients with hemophilia.

Journal Article Gene Ther · March 2012 Recombinant adeno-associated virus (rAAV) is a promising gene delivery vector and has recently been used in patients with hemophilia. One limitation of AAV application is that most humans have experienced wild-type AAV serotype 2 exposure, which frequently ... Full text Link to item Cite

Intra- and inter-subunit disulfide bond formation is nonessential in adeno-associated viral capsids.

Journal Article PLoS One · 2012 The capsid proteins of adeno-associated viruses (AAV) have five conserved cysteine residues. Structural analysis of AAV serotype 2 reveals that Cys289 and Cys361 are located adjacent to each other within each monomer, while Cys230 and Cys394 are located on ... Full text Link to item Cite

Production of recombinant adeno-associated viral vectors and use in in vitro and in vivo administration.

Journal Article Curr Protoc Neurosci · October 2011 Adeno-associated virus is a nonpathogenic human virus that has been developed into a gene-delivery vector due to its high efficiency of infection for many different cell types and its ability to persist and lead to long-term gene expression. This unit desc ... Full text Link to item Cite

Peptide affinity reagents for AAV capsid recognition and purification.

Journal Article Gene Ther · October 2011 We report the discovery of AAV capsid-binding peptides identified through phage panning. The heptapeptide motif GYVSRHP selectively recognized AAV serotype 8 capsids and blocked transduction in vitro. Recombinant AAV8 vectors were purified directly from cr ... Full text Link to item Cite

Engineering liver-detargeted AAV9 vectors for cardiac and musculoskeletal gene transfer.

Journal Article Mol Ther · June 2011 We report the generation of a new class of adeno-associated virus serotype 9 (AAV9)-derived vectors displaying selective loss of liver tropism and demonstrating potential for cardiac and musculoskeletal gene transfer applications. Random mutagenesis of res ... Full text Link to item Cite

Glycated AAV vectors: chemical redirection of viral tissue tropism.

Journal Article Bioconjug Chem · April 20, 2011 A chemical approach for selective masking of arginine residues on viral capsids featuring an exogenous glycation reaction has been developed. Reaction of adeno-associated viral (AAV) capsids with the α-dicarbonyl compound, methylglyoxal, resulted in format ... Full text Link to item Cite

Terminal N-linked galactose is the primary receptor for adeno-associated virus 9.

Journal Article J Biol Chem · April 15, 2011 Sialylated glycans serve as cell surface attachment factors for a broad range of pathogens. We report an atypical example, where desialylation increases cell surface binding and infectivity of adeno-associated virus (AAV) serotype 9, a human parvovirus iso ... Full text Link to item Cite

Systemic gene transfer to skeletal muscle using reengineered AAV vectors.

Journal Article Methods Mol Biol · 2011 Gene therapy of musculoskeletal disorders warrants efficient gene transfer to a wide range of muscle groups. Reengineered adeno-associated viral (AAV) vectors that selectively transduce muscle tissue following systemic administration are attractive candida ... Full text Link to item Cite

Bocavirus episome in infected human tissue contains non-identical termini.

Journal Article PLoS One · 2011 UNLABELLED: Human bocaviruses (HBoV) are highly prevalent human infections whose pathogenic potential remains unknown. Recent identification of the first non-human primate bocavirus [1] in captive gorillas raised the possibility of the persistent nature of ... Full text Link to item Cite

Reengineered AAV vectors: old dog, new tricks.

Journal Article Discov Med · May 2010 Adeno-associated viral (AAV) vectors have emerged in recent years as powerful tools for therapeutic gene transfer. Successes in clinical trials and the discovery of several hundreds of naturally occurring AAV isolates have triggered efforts to understand a ... Link to item Cite

Reengineering a receptor footprint of adeno-associated virus enables selective and systemic gene transfer to muscle.

Journal Article Nat Biotechnol · January 2010 Reengineering the receptor footprints of adeno-associated virus (AAV) isolates may yield variants with improved properties for clinical applications. We generated a panel of synthetic AAV2 vectors by replacing a hexapeptide sequence in a previously identif ... Full text Link to item Cite

Cytotoxic-T-lymphocyte-mediated elimination of target cells transduced with engineered adeno-associated virus type 2 vector in vivo.

Journal Article J Virol · July 2009 A recent clinical trial in patients with hemophilia B has suggested that adeno-associated virus (AAV) capsid-specific cytotoxic T lymphocytes (CTLs) eliminated AAV-transduced hepatocytes and resulted in therapeutic failure. AAV capsids elicit a CTL respons ... Full text Link to item Cite

Cellular immune response to cryptic epitopes during therapeutic gene transfer.

Journal Article Proc Natl Acad Sci U S A · June 30, 2009 The immune response has been implicated as a critical factor in determining the success or failure of clinical gene therapy trials. Generally, such a response is elicited by the desired transgene product or, in some cases, the delivery system. In the curre ... Full text Link to item Cite

A small regulatory element from chromosome 19 enhances liver-specific gene expression.

Journal Article Gene Ther · January 2009 Tissue-specific promoters for gene therapy are typically too big for adeno-associated virus (AAV) vectors; thus, the exploration of small effective non-viral regulatory elements is of particular interest. Wild-type AAV can specifically integrate into a reg ... Full text Link to item Cite

Bioluminescent virion shells: new tools for quantitation of AAV vector dynamics in cells and live animals.

Journal Article Gene Ther · December 2008 Current technologies for visualizing infectious pathways of viruses rely on fluorescent labeling of capsid proteins by chemical conjugation or genetic manipulation. For noninvasive in vivo imaging of such agents in mammalian tissue, we engineered biolumine ... Full text Link to item Cite

Engineering and Selection of Shuffled AAV Genomes: A New Strategy for Producing Targeted Biological Nanoparticles.

Journal Article Mol Ther · July 2008 We report a DNA shuffling-based approach for developing cell type-specific vectors through directed evolution. Capsid genomes of adeno-associated virus (AAV) serotypes 1-9 were randomly fragmented and reassembled using PCR to generate a chimeric capsid lib ... Full text Link to item Cite

Surface loop dynamics in adeno-associated virus capsid assembly.

Journal Article J Virol · June 2008 The HI loop is a prominent domain on the adeno-associated virus (AAV) capsid surface that extends from each viral protein (VP) subunit overlapping the neighboring fivefold VP. Despite the highly conserved nature of the residues at the fivefold pore, the HI ... Full text Link to item Cite

Adeno-associated virus type 2 (AAV2) capsid-specific cytotoxic T lymphocytes eliminate only vector-transduced cells coexpressing the AAV2 capsid in vivo.

Journal Article J Virol · July 2007 A recent clinical trial has suggested that recombinant adeno-associated virus (rAAV) vector transduction in humans induces a cytotoxic T-lymphocyte (CTL) response against the AAV2 capsid. To directly address the ability of AAV capsid-specific CTLs to elimi ... Full text Link to item Cite

Production of recombinant adeno-associated viral vectors for in vitro and in vivo use.

Journal Article Curr Protoc Mol Biol · April 2007 Adeno-associated virus is a nonpathogenic human virus that has been developed into a gene-delivery vector due to its high efficiency of infection in many different cell types and its ability to persist and lead to long-term gene expression. The vector is a ... Full text Link to item Cite

Production of recombinant adeno-associated viral vectors.

Journal Article Curr Protoc Hum Genet · April 2007 Adeno-associated virus is a nonpathogenic human virus that has been developed into a gene-delivery vector due to its high efficiency of infection for many different cell types and its ability to persist and lead to long-term gene expression. This unit desc ... Full text Link to item Cite

Single amino acid changes can influence titer, heparin binding, and tissue tropism in different adeno-associated virus serotypes.

Journal Article J Virol · November 2006 Despite the high degree of sequence homology between adeno-associated virus (AAV) serotype 1 and 6 capsids (99.2%), these viruses have different liver transduction profiles when tested as vectors. Examination of the six amino acid residues that differ betw ... Full text Link to item Cite

Adeno-associated virus type 2 contains an integrin alpha5beta1 binding domain essential for viral cell entry.

Journal Article J Virol · September 2006 Integrins have been implicated as coreceptors in the infectious pathways of several nonenveloped viruses. For example, adenoviruses are known to interact with alphaV integrins by virtue of a high-affinity arginine-glycine-aspartate (RGD) domain present in ... Full text Link to item Cite

Adeno-associated virus serotypes: vector toolkit for human gene therapy.

Journal Article Mol Ther · September 2006 Recombinant adeno-associated viral (AAV) vectors have rapidly advanced to the forefront of gene therapy in the past decade. The exponential progress of AAV-based vectors has been made possible by the isolation of several naturally occurring AAV serotypes a ... Full text Link to item Cite

Production of recombinant adeno-associated viral vectors and use for in vitro and in vivo administration.

Journal Article Curr Protoc Neurosci · May 2006 Adeno-associated virus is a nonpathogenic human virus that has been developed into a gene-delivery vector due to its high efficiency of infection for many different cell types and its ability to persist and lead to long-term gene expression. This unit desc ... Full text Link to item Cite

AAV does the shuffle.

Journal Article Nat Biotechnol · February 2006 Full text Link to item Cite

Cytosolic delivery of macromolecules 4. Head group-dependent membrane permeabilization by pH-sensitive detergents.

Journal Article J Control Release · August 18, 2005 Three tertiary amine-based detergents with zero, one, or two hydroxyl groups at various positions in their head group were characterized for their ability to promote the cytosolic delivery of macromolecules. Critical micellar concentrations (CMC) and membr ... Full text Link to item Cite

Cytosolic delivery of macromolecules. 3. Synthesis and characterization of acid-sensitive bis-detergents.

Conference Bioconjug Chem · 2004 A serious limitation that precludes utilization of single-tailed, pH-sensitive detergents for the cytosolic delivery of macromolecules is their low limit of incorporation in stable liposomal formulations. To address this issue, we have prepared two Gemini ... Full text Link to item Cite

Cytosolic delivery of macromolecules. II. Mechanistic studies with pH-sensitive morpholine lipids.

Journal Article Biochim Biophys Acta · April 1, 2003 Drug carriers containing weak acids or bases can promote cytosolic delivery of macromolecules by exploiting the acidic pH of the endosome. We have prepared two pH-sensitive mono-stearoyl derivatives of morpholine, one with a (2-hydroxy) propylene (ML1) lin ... Full text Link to item Cite

Cytosolic delivery of macromolecules: I. Synthesis and characterization of pH-sensitive acyloxyalkylimidazoles.

Journal Article Biochim Biophys Acta · April 1, 2003 A series of 1-(acyloxyalkyl)imidazoles (AAI) were synthesized by nucleophilic substitution of chloroalkyl esters of fatty acids with imidazole. The former was prepared from fatty acid chloride and an aldehyde. When incorporated into liposomes, these lipids ... Full text Link to item Cite

Exploitation of intracellular pH gradients in the cellular delivery of macromolecules.

Journal Article J Pharm Sci · April 2002 Most cellular components such as the cytoplasm, endosomes, lysosomes, endoplasmic reticulum, Golgi bodies, mitochondria, and nuclei are known to maintain their own characteristic pH values. These pH values range from as low as 4.5 in the lysosome to about ... Full text Link to item Cite