Work

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

Year: 2024

Type: article

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. • Genome editing of swine cells using the CRISPR-Cas9 system • Description of in vitro cleavage assay to test RNP complex functionality • Fast and cost-effective method for assessing on-target editing efficiency Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2024.103385