Generation of Large Fragment Knock-In Mouse Models by Microinjecting into 2-Cell Stage Embryos

Methods Mol Biol. 2020:2066:89-100. doi: 10.1007/978-1-4939-9837-1_7.

Abstract

Large fragment knock-in mouse models such as reporters and conditional mutant mice are important models for biological research. Here we describe 2-cell (2C)-homologous recombination (HR)-CRISPR, a highly efficient method to generate large fragment knock-in mouse models by CRISPR-based genome engineering. Using this method, knock-in founders can be generated routinely in a time frame of about two months with high germline transmission efficiency. 2C-HR-CRISPR will significantly promote the advancement of basic and translational research using genetic mouse models.

Keywords: 2-Cell stage; CRISPR-Cas9; Homologous recombination; Knock-in.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics
  • Embryo, Mammalian
  • Embryonic Development / genetics*
  • Gene Editing / methods
  • Gene Knock-In Techniques / methods*
  • Genome / genetics*
  • Homologous Recombination / genetics
  • Mice
  • Microinjections / methods*

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