CRISPR mediated somatic cell genome engineering in the chicken

Dev Biol. 2015 Nov 1;407(1):68-74. doi: 10.1016/j.ydbio.2015.08.007. Epub 2015 Aug 13.

Abstract

Gene-targeted knockout technologies are invaluable tools for understanding the functions of genes in vivo. CRISPR/Cas9 system of RNA-guided genome editing is revolutionizing genetics research in a wide spectrum of organisms. Here, we combined CRISPR with in vivo electroporation in the chicken embryo to efficiently target the transcription factor PAX7 in tissues of the developing embryo. This approach generated mosaic genetic mutations within a wild-type cellular background. This series of proof-of-principle experiments indicate that in vivo CRISPR-mediated cell genome engineering is an effective method to achieve gene loss-of-function in the tissues of the chicken embryo and it completes the growing genetic toolbox to study the molecular mechanisms regulating development in this important animal model.

Keywords: CRISPR; Cas9; Chicken; Electroporation; Gene-targeted knockout; PAX7.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Chick Embryo
  • Chickens
  • Clustered Regularly Interspaced Short Palindromic Repeats / physiology*
  • Electroporation
  • Genetic Engineering*
  • Genome
  • Molecular Sequence Data
  • PAX7 Transcription Factor / physiology

Substances

  • PAX7 Transcription Factor