Generation of Large Fragment Knock-In Mouse Models by Microinjecting into 2-Cell Stage Embryos.
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.
['Gu|Bin|B|', 'Gertsenstein|Marina|M|', 'Posfai|Eszter|E|']
D000818:Animals D064113:CRISPR-Cas Systems D004622:Embryo, Mammalian D047108:Embryonic Development D000072669:Gene Editing D055879:Gene Knock-In Techniques D016678:Genome D059765:Homologous Recombination D051379:Mice D008845:Microinjections
Knock-in 2-Cell stage CRISPR-Cas9 Homologous recombination