Electroporation-Mediated CRISPR/Cas9 Genome Editing in Rat Zygotes

Daniel J Davis; Hongsheng Men; Elizabeth C Bryda

doi:10.1007/978-1-0716-2990-1_11

Electroporation-Mediated CRISPR/Cas9 Genome Editing in Rat Zygotes

Methods Mol Biol. 2023:2631:267-276. doi: 10.1007/978-1-0716-2990-1_11.

Authors

Daniel J Davis¹, Hongsheng Men², Elizabeth C Bryda^{3

2}

Affiliations

¹ Animal Modeling Core, University of Missouri, Columbia, MO, USA. davisdaniel@missouri.edu.
² Rat Resource and Research Center, University of Missouri, Columbia, MO, USA.
³ Animal Modeling Core, University of Missouri, Columbia, MO, USA.

PMID: 36995672
DOI: 10.1007/978-1-0716-2990-1_11

Abstract

Genetic engineering in the rat has been revolutionized by the development of CRISPR-based genome editing tools. Conventional methods for inserting genome editing elements such as CRISPR/Cas9 reagents into rat zygotes include cytoplasmic or pronuclear microinjections. These techniques are labor-intensive, require specialized micromanipulator equipment, and are technically challenging. Here, we describe a simple and effective method for zygote electroporation in which CRISPR/Cas9 reagents are introduced into rat zygotes via pores produced by precise electrical pulses applied to the cells. Zygote electroporation allows for high-throughput efficient genome editing in rat embryos.

Keywords: CRISPR/Cas9; Electroporation; Embryo; Genome editing; Rat; Zygote.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Cas Systems* / genetics
Electroporation / methods
Electroporation Therapies
Gene Editing* / methods
Rats
Zygote