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Methods Mol Biol · 2022

AAV-Mediated Somatic Gene Editing for Cardiac and Skeletal Muscle in a Large Animal Model.

Methods in molecular biology (Clifton, N.J.), 2573: 63-74 PMID 36040587 DOI 10.1007/978-1-0716-2707-5_6

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Abstract

Here we describe a protocol to produce a recombinant adeno-associated viral vector (rAAV)-based system to deliver the CRISPR-Cas9 complex into porcine skeletal muscle and myocardial cells. We initially describe the genomic composition of the rAAV-CRISPR vectors used in our lab. Furthermore, we give a step-by-step instruction into the production of recombinant viral vectors with high yields and purity. Lastly we describe the minimally invasive injection regimes to target the myocardium in a pig.

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