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

Bin Gu, Marina Gertsenstein, Eszter Posfai

Research output: Chapter in Book/Report/Conference proceedingChapter

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)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages89-100
Number of pages12
DOIs
StatePublished - Jan 1 2020

Publication series

NameMethods in Molecular Biology
Volume2066

Fingerprint

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

Gu, B., Gertsenstein, M., & Posfai, E. (2020). Generation of Large Fragment Knock-In Mouse Models by Microinjecting into 2-Cell Stage Embryos. In Methods in Molecular Biology (pp. 89-100). (Methods in Molecular Biology; Vol. 2066). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9837-1_7
Gu, Bin ; Gertsenstein, Marina ; Posfai, Eszter. / Generation of Large Fragment Knock-In Mouse Models by Microinjecting into 2-Cell Stage Embryos. Methods in Molecular Biology. Humana Press Inc., 2020. pp. 89-100 (Methods in Molecular Biology).
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Gu, B, Gertsenstein, M & Posfai, E 2020, Generation of Large Fragment Knock-In Mouse Models by Microinjecting into 2-Cell Stage Embryos. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 2066, Humana Press Inc., pp. 89-100. https://doi.org/10.1007/978-1-4939-9837-1_7

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

Methods in Molecular Biology. Humana Press Inc., 2020. p. 89-100 (Methods in Molecular Biology; Vol. 2066).

Research output: Chapter in Book/Report/Conference proceedingChapter

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Gu B, Gertsenstein M, Posfai E. Generation of Large Fragment Knock-In Mouse Models by Microinjecting into 2-Cell Stage Embryos. In Methods in Molecular Biology. Humana Press Inc. 2020. p. 89-100. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9837-1_7