清脆的
基因组编辑
合子
电穿孔
Cas9
斑马鱼
生物
计算生物学
基因组
细胞生物学
遗传学
胚胎
基因
胚胎发生
作者
Daniel J. Davis,Hongsheng Men,Elizabeth C. Bryda
出处
期刊:Methods in molecular biology
日期:2023-01-01
卷期号:: 267-276
被引量:1
标识
DOI:10.1007/978-1-0716-2990-1_11
摘要
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.
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