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

Publication ,  Journal Article
Li, W; Asokan, A; Wu, Z; Van Dyke, T; DiPrimio, N; Johnson, JS; Govindaswamy, L; Agbandje-McKenna, M; Leichtle, S; Eugene Redmond, D ...
Published in: 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 library. A single infectious clone (chimeric-1829) containing genome fragments from AAV1, 2, 8, and 9 was isolated from an integrin minus hamster melanoma cell line previously shown to have low permissiveness to AAV. Molecular modeling studies suggest that AAV2 contributes to surface loops at the icosahedral threefold axis of symmetry, while AAV1 and 9 contribute to two- and fivefold symmetry interactions, respectively. The C-terminal domain (AAV9) was identified as a critical structural determinant of melanoma tropism through rational mutagenesis. Chimeric-1829 utilizes heparan sulfate as a primary receptor and transduces melanoma cells more efficiently than all serotypes. Further, chimeric-1829 demonstrates altered tropism in rodent skeletal muscle, liver, and brain including nonhuman primates. We determined a unique immunological profile based on neutralizing antibody (NAb) titer and crossreactivity studies strongly supporting isolation of a synthetic laboratory-derived capsid variant. Application of this technology to alternative cell/tissue types using AAV or other viral capsid sequences is likely to yield a new class of biological nanoparticles as vectors for human gene transfer.

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Published In

Mol Ther

DOI

EISSN

1525-0024

Publication Date

July 2008

Volume

16

Issue

7

Start / End Page

1252 / 1260

Location

United States

Related Subject Headings

  • Virus Internalization
  • Transduction, Genetic
  • Primates
  • Nanoparticles
  • Muscle, Skeletal
  • Mice, Inbred BALB C
  • Mice
  • Melanoma
  • Liver
  • Humans
 

Citation

APA
Chicago
ICMJE
MLA
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Li, W., Asokan, A., Wu, Z., Van Dyke, T., DiPrimio, N., Johnson, J. S., … Samulski, R. J. (2008). Engineering and Selection of Shuffled AAV Genomes: A New Strategy for Producing Targeted Biological Nanoparticles. Mol Ther, 16(7), 1252–1260. https://doi.org/10.1038/mt.2008.100
Li, Wuping, Aravind Asokan, Zhijian Wu, Terry Van Dyke, Nina DiPrimio, Jarrod S. Johnson, Lakshmanan Govindaswamy, et al. “Engineering and Selection of Shuffled AAV Genomes: A New Strategy for Producing Targeted Biological Nanoparticles.Mol Ther 16, no. 7 (July 2008): 1252–60. https://doi.org/10.1038/mt.2008.100.
Li W, Asokan A, Wu Z, Van Dyke T, DiPrimio N, Johnson JS, et al. Engineering and Selection of Shuffled AAV Genomes: A New Strategy for Producing Targeted Biological Nanoparticles. Mol Ther. 2008 Jul;16(7):1252–60.
Li, Wuping, et al. “Engineering and Selection of Shuffled AAV Genomes: A New Strategy for Producing Targeted Biological Nanoparticles.Mol Ther, vol. 16, no. 7, July 2008, pp. 1252–60. Pubmed, doi:10.1038/mt.2008.100.
Li W, Asokan A, Wu Z, Van Dyke T, DiPrimio N, Johnson JS, Govindaswamy L, Agbandje-McKenna M, Leichtle S, Eugene Redmond D, McCown TJ, Petermann KB, Sharpless NE, Samulski RJ. Engineering and Selection of Shuffled AAV Genomes: A New Strategy for Producing Targeted Biological Nanoparticles. Mol Ther. 2008 Jul;16(7):1252–1260.

Published In

Mol Ther

DOI

EISSN

1525-0024

Publication Date

July 2008

Volume

16

Issue

7

Start / End Page

1252 / 1260

Location

United States

Related Subject Headings

  • Virus Internalization
  • Transduction, Genetic
  • Primates
  • Nanoparticles
  • Muscle, Skeletal
  • Mice, Inbred BALB C
  • Mice
  • Melanoma
  • Liver
  • Humans