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

Bin Gu, Marina Gertsenstein, Eszter Posfai

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)89-100
Number of pages12
JournalMethods in molecular biology (Clifton, N.J.)
Volume2066
DOIs
StatePublished - Jan 1 2020

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Clustered Regularly Interspaced Short Palindromic Repeats
Embryonic Structures
Homologous Recombination
Biological Models
Translational Medical Research
Genetic Models
Genome
Research

All Science Journal Classification (ASJC) codes

  • Genetics
  • Molecular Biology

Keywords

  • 2-Cell stage
  • CRISPR-Cas9
  • Homologous recombination
  • Knock-in

Cite this

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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.",
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Generation of Large Fragment Knock-In Mouse Models by Microinjecting into 2-Cell Stage Embryos. / Gu, Bin; Gertsenstein, Marina; Posfai, Eszter.

In: Methods in molecular biology (Clifton, N.J.), Vol. 2066, 01.01.2020, p. 89-100.

Research output: Contribution to journalArticle

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