Davis Daniel J, Men Hongsheng, Bryda Elizabeth C
Animal Modeling Core, University of Missouri, Columbia, MO, USA.
Rat Resource and Research Center, University of Missouri, Columbia, MO, USA.
Methods Mol Biol. 2023;2631:267-276. doi: 10.1007/978-1-0716-2990-1_11.
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.
基于CRISPR的基因组编辑工具的发展彻底改变了大鼠的基因工程。将基因组编辑元件（如CRISPR/Cas9试剂）插入大鼠受精卵的传统方法包括细胞质或原核显微注射。这些技术劳动强度大，需要专门的显微操作设备，并且在技术上具有挑战性。在这里，我们描述了一种简单有效的受精卵电穿孔方法，其中通过对细胞施加精确的电脉冲产生的孔将CRISPR/Cas9试剂引入大鼠受精卵。受精卵电穿孔可实现大鼠胚胎的高通量高效基因组编辑。
