Generation of genetically modified rat models via the CRISPR/Cas9 technology

Mei Zhen Liu, Li Ren Wang, Yong Mei Li, Xue Yun Ma, Hong Hui Han, Da Li Li

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

RNA介导的CRISPR/Cas9基因编辑系统由单链引导RNA(sgRNA)与核酸酶Cas9构成。在细胞内,sgRNA能够按照碱基互补配对的原则引导Cas9与靶点结合,由Cas9切割目标DNA,造成双链DNA断裂(double stranded break, DSB)。在随后的DNA修复过程中,细胞主要进行非同源末端连接(non-homologous end joining, NHEJ)或在有修复模板存在的情况下进行重组修复(homology directed repair, HDR)。如果将CRISPR/Cas9系统以及修复模板通过显微注射的方式导入大鼠的胚胎内,就能借助细胞的修复机制实现大鼠胚胎的基因编辑,由此构建各种基因修饰大鼠模型。本文详细介绍了利用CRISPR/Cas9基因编辑技术构建大鼠模型的具体操作步骤,以期为相关领域的科研人员提供一种大鼠基因修饰模型的构建方法。.

Original languageEnglish
Pages (from-to)78-87
Number of pages10
JournalYi chuan = Hereditas / Zhongguo yi chuan xue hui bian ji
Volume45
Issue number1
DOIs
StatePublished - 20 Jan 2023

Keywords

  • CRISPR/Cas9
  • gene knockout
  • genome editing
  • rat

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