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

Patricia Kiesler; Stella S. Lee; Alexis L. Norris; Mayumi F. Miller; Carlo J. Mercado; Adam L. Moyer; Samantha Maragh

STAR Protocols (Dec 2024)

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

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

Affiliations

Patricia Kiesler: Biosystems and Biomaterials Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA; Corresponding author
Stella S. Lee: Center for Veterinary Medicine, Office of New Animal Drug Evaluation, US Food and Drug Administration, Rockville, MD 20855, USA
Alexis L. Norris: Center for Veterinary Medicine, Office of New Animal Drug Evaluation, US Food and Drug Administration, Rockville, MD 20855, USA
Mayumi F. Miller: Center for Veterinary Medicine, Office of Applied Science, US Food and Drug Administration, Laurel, MD 20708, USA
Carlo J. Mercado: Center for Veterinary Medicine, Office of New Animal Drug Evaluation, US Food and Drug Administration, Rockville, MD 20855, USA
Adam L. Moyer: Center for Veterinary Medicine, Office of New Animal Drug Evaluation, US Food and Drug Administration, Rockville, MD 20855, USA
Samantha Maragh: Biosystems and Biomaterials Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA; Corresponding author

Journal volume & issue: Vol. 5, no. 4
p. 103385

Abstract

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Summary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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