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

Electroporation-Mediated CRISPR/Cas9 Genome Editing in Rat Zygotes

Daniel J Davis  1 , Hongsheng Men  2 , Elizabeth C Bryda  3   2
Affiliations

Electroporation-Mediated CRISPR/Cas9 Genome Editing in Rat Zygotes

Daniel J Davis et al. Methods Mol Biol. 2023.

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.

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