Protocol for CRISPR-Cas9 genome editing of a swine cell line via electroporation
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.
Authors
- Alexis L. Norris (ORCID: https://orcid.org/0000-0002-0442-9096)
- Adam L. Moyer (ORCID: https://orcid.org/0000-0001-7802-2777)
- Stella S. Lee (ORCID: https://orcid.org/0000-0002-2629-2422)
- Patricia Kiesler
- Carlo J. Mercado
- Samantha Maragh (ORCID: https://orcid.org/0000-0003-2564-9589)
- Mayumi F. Miller (ORCID: https://orcid.org/0000-0002-3794-7463)
Institutions
- Center for Veterinary Medicine (US)
- National Institute of Standards and Technology (US)
Publication Details
- Journal
- STAR Protocols
- Published
- 2024-10-10
- DOI
- https://doi.org/10.1016/j.xpro.2024.103385
- Primary Topic
- CRISPR and Genetic Engineering
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- U.S. Food and Drug Administration
- National Institute of Standards and Technology