Protocol for CRISPR-Cas9 genome editing of a swine cell line via electroporation

利用电穿孔法对猪细胞系进行CRISPR-Cas9基因组编辑的实验方案

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作者:Patricia Kiesler ,Stella S Lee ,Alexis L Norris ,Mayumi F Miller ,Carlo J Mercado ,Adam L Moyer ,Samantha Maragh

Abstract

Genome editing technology is being used in animals for a variety of purposes, including improvement of animal and public health outcomes. Characterization of genome editing reagents and anticipated genomic alterations is an essential step toward the development of an edited animal. Here, we present a protocol for genome editing in the swine testicular (ST) cell line. We describe steps for evaluating CRISPR-Cas9 complex functionality in vitro, delivering editing molecules into cells by transfection, and assessing target editing via Sanger sequencing. Keywords: Biotechnology and bioengineering; CRISPR; Cell-based Assays.

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