Journal
/
/
CRISPR/Cas9-Mediated Highly Efficient Gene Targeting in Embryonic Stem Cells for Developing Gene-Manipulated Mouse Models
Journal JoVE
Biologie du développement
This content is Free Access.
Journal JoVE Biologie du développement
CRISPR/Cas9-Mediated Highly Efficient Gene Targeting in Embryonic Stem Cells for Developing Gene-Manipulated Mouse Models
DOI:

10:57 min

August 24, 2022

, ,

Chapitres

  • 00:04Introduction
  • 01:06Gene Targeting of Embryonic Stem Cells (ESCs)
  • 04:16PCR Genotyping of Targeted ESCs
  • 07:02Preparation of Eight-Cell Stage Embryo and Microinjection of ESCs
  • 09:49Results: Screening of Targeted ESC Clones
  • 10:10Conclusion

Summary

Traduction automatique

Here we present a protocol for developing genetically modified mouse models using embryonic stem cells, especially for large DNA knock-in (KI). This protocol is tuned up using CRISPR/Cas9 genome editing, resulting in significantly improved KI efficiency compared with the conventional homologous recombination-mediated linearized DNA targeting method.

Vidéos Connexes

Read Article