Skip to main content

Somatic Liver Knockout (SLiK): A Quick and Efficient Way to Generate Liver-Specific Knockout Mice Using Multiplex CRISPR/Cas9 Gene Editing.

Publication ,  Journal Article
Johnson, CG; Chen, T; Furey, N; Hemmingsen, MG; Bissig, K-D
Published in: Current protocols in molecular biology
March 2020

Somatic liver knockout (SLiK) is a method developed to rapidly generate a liver-specific knockout of one or several genes. This technique combines the strengths of CRISPR/Cas9 gene editing and hydrodynamic tail-vein injection, a simple in vivo method for transfection of hepatocytes, to harness the powerful selection pressure of tyrosinemic livers to replace host hepatocytes with any desired gene deletion. In this protocol, we will describe sgRNA design and cloning, hydrodynamic tail-vein injection of targeting constructs, and screening and validation methods for efficient in vivo gene editing. © 2020 by John Wiley & Sons, Inc. Support Protocol 1: sgRNA design Support Protocol 2: sgRNA construction: daisy chaining multiple sgRNAs Basic Protocol: Delivery of DNA by hydrodynamic tail-vein injection and liver repopulation of edited hepatocytes Support Protocol 3: Validation of CRISPR/Cas9 cutting in vivo.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

Current protocols in molecular biology

DOI

EISSN

1934-3647

ISSN

1934-3639

Publication Date

March 2020

Volume

130

Issue

1

Start / End Page

e117

Related Subject Headings

  • Transfection
  • RNA, Guide, CRISPR-Cas Systems
  • Mice, Knockout
  • Mice
  • Liver
  • Hepatocytes
  • Gene Editing
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • CRISPR-Cas Systems
  • Biochemistry & Molecular Biology
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Johnson, C. G., Chen, T., Furey, N., Hemmingsen, M. G., & Bissig, K.-D. (2020). Somatic Liver Knockout (SLiK): A Quick and Efficient Way to Generate Liver-Specific Knockout Mice Using Multiplex CRISPR/Cas9 Gene Editing. Current Protocols in Molecular Biology, 130(1), e117. https://doi.org/10.1002/cpmb.117
Johnson, Collin G., Tong Chen, Nika Furey, Madeline G. Hemmingsen, and Karl-Dimiter Bissig. “Somatic Liver Knockout (SLiK): A Quick and Efficient Way to Generate Liver-Specific Knockout Mice Using Multiplex CRISPR/Cas9 Gene Editing.Current Protocols in Molecular Biology 130, no. 1 (March 2020): e117. https://doi.org/10.1002/cpmb.117.
Johnson CG, Chen T, Furey N, Hemmingsen MG, Bissig K-D. Somatic Liver Knockout (SLiK): A Quick and Efficient Way to Generate Liver-Specific Knockout Mice Using Multiplex CRISPR/Cas9 Gene Editing. Current protocols in molecular biology. 2020 Mar;130(1):e117.
Johnson, Collin G., et al. “Somatic Liver Knockout (SLiK): A Quick and Efficient Way to Generate Liver-Specific Knockout Mice Using Multiplex CRISPR/Cas9 Gene Editing.Current Protocols in Molecular Biology, vol. 130, no. 1, Mar. 2020, p. e117. Epmc, doi:10.1002/cpmb.117.
Johnson CG, Chen T, Furey N, Hemmingsen MG, Bissig K-D. Somatic Liver Knockout (SLiK): A Quick and Efficient Way to Generate Liver-Specific Knockout Mice Using Multiplex CRISPR/Cas9 Gene Editing. Current protocols in molecular biology. 2020 Mar;130(1):e117.

Published In

Current protocols in molecular biology

DOI

EISSN

1934-3647

ISSN

1934-3639

Publication Date

March 2020

Volume

130

Issue

1

Start / End Page

e117

Related Subject Headings

  • Transfection
  • RNA, Guide, CRISPR-Cas Systems
  • Mice, Knockout
  • Mice
  • Liver
  • Hepatocytes
  • Gene Editing
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • CRISPR-Cas Systems
  • Biochemistry & Molecular Biology