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. 2019:1950:123-139.
doi: 10.1007/978-1-4939-9139-6_7.

Use of AAV Vectors for CRISPR-Mediated In Vivo Genome Editing in the Retina

Wenhan Yu  1 , Zhijian Wu  2
Affiliations

Use of AAV Vectors for CRISPR-Mediated In Vivo Genome Editing in the Retina

Wenhan Yu et al. Methods Mol Biol. 2019.

Abstract

Degenerative retinal diseases such as retinitis pigmentosa (RP) and Leber's congenital amaurosis (LCA) may lead to blindness without effective treatment. With the rapid advancement of the CRISPR/Cas9 genome editing technology, in vivo application of CRISPR/Cas9 holds immense potential for treatment of these diseases. Adeno-associated virus (AAV) vectors are an ideal gene transfer tool for delivery of CRISPR components to the retina. Here, we describe a protocol for utilizing an AAV-based CRISPR/Cas9 system for in vivo genome editing in the retina.

Keywords: AAV vector; CRISPR/Cas9; Gene therapy; Genome editing; Retina; Retinal degeneration.

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Figures

Fig. 1
Fig. 1
Sample SURVEYOR assay result. The SURVEYOR nuclease recognizes the mismatch (if any) in the reannealed heteroduplex, which results from indels introduced into the predicted cleavage site in the CRISPR edited cells. Black arrow: undigested amplicons; black triangles: cleavage products of the SURVEYOR nuclease
See this image and copyright information in PMC

References

    1. Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, Norville JE, Church GM (2013) RNA-guided human genome engineering via Cas9. Science 339(6121):823–826. 10.1126/science.1232033 - DOI - PMC - PubMed
    1. Lau CH, Suh Y (2017) In vivo genome editing in animals using AAV-CRISPR system: applications to translational research of human disease. F1000Res 6:2153 10.12688/f1000research.11243.1 - DOI - PMC - PubMed
    1. Wang S, Sengel C, Emerson MM, Cepko CL (2014) A gene regulatory network controls the binary fate decision of rod and bipolar cells in the vertebrate retina. Dev Cell 30(5):513–527. 10.1016/j.devcel.2014.07.018 - DOI - PMC - PubMed
    1. Bakondi B, Lv W, Lu B, Jones MK, Tsai Y, Kim KJ, Levy R, Akhtar AA, Breunig JJ, Svendsen CN, Wang S (2016) In vivo CRISPR/Cas9 gene editing corrects retinal dystrophy in the S334ter-3 rat model of autosomal dominant retinitis pigmentosa. Mol Ther 24 (3):556–563. 10.1038/mt.2015.220 - DOI - PMC - PubMed
    1. Latella MC, Di Salvo MT, Cocchiarella F, Benati D, Grisendi G, Comitato A, Marigo V, Recchia A (2016) In vivo editing of the human mutant rhodopsin gene by electroporation of plasmid-based CRISPR/Cas9 in the mouse retina. Mol Ther Nucleic Acids 5(11):e389 10.1038/mtna.2016.92 - DOI - PMC - PubMed

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