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

STAR Protoc. 2024 Dec 20;5(4):103385. doi: 10.1016/j.xpro.2024.103385. Epub 2024 Oct 10.

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.

MeSH terms

  • Animals
  • CRISPR-Cas Systems* / genetics
  • Cell Line
  • Electroporation* / methods
  • Gene Editing* / methods
  • Male
  • Swine
  • Testis / cytology
  • Testis / metabolism
  • Transfection / methods