Generation of genetically modified mice through the microinjection of oocytes

Fabien Delerue*, Lars M. Ittner

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)

Abstract

The use of genetically modified mice has significantly contributed to studies on both physiological and pathological in vivo processes. The pronuclear injection of DNA expression constructs into fertilized oocytes remains the most commonly used technique to generate transgenic mice for overexpression. With the introduction of CRISPR technology for gene targeting, pronuclear injection into fertilized oocytes has been extended to the generation of both knockout and knockin mice. This work describes the preparation of DNA for injection and the generation of CRISPR guides for gene targeting, with a particular emphasis on quality control. The genotyping procedures required for the identification of potential founders are critical. Innovative genotyping strategies that take advantage of the “multiplexing” capabilities of CRISPR are presented herein. Surgical procedures are also outlined. Together, the steps of the protocol will allow for the generation of genetically modified mice and for the subsequent establishment of mouse colonies for a plethora of research fields, including immunology, neuroscience, cancer, physiology, development, and others.

Original languageEnglish
Article numbere55765
Pages (from-to)1-12
Number of pages12
JournalJournal of Visualized Experiments
Volume2017
Issue number124
DOIs
Publication statusPublished - 15 Jun 2017
Externally publishedYes

Keywords

  • CRISPR
  • Cytoplasmic
  • Developmental Biology
  • Genome editing
  • Issue 124
  • Mice
  • Microinjection
  • Oocytes
  • Pronuclear
  • Transgenic

Fingerprint

Dive into the research topics of 'Generation of genetically modified mice through the microinjection of oocytes'. Together they form a unique fingerprint.

Cite this