CRISPR mediated somatic cell genome engineering in the chicken

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...

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Bibliographic Details
Published in:Developmental biology 2015-11, Vol.407 (1), p.68-74
Main Authors: Véron, Nadège, Qu, Zhengdong, Kipen, Phoebe A.S., Hirst, Claire E., Marcelle, Christophe
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Cas9
Chick Embryo
Chicken
Chickens
Clustered Regularly Interspaced Short Palindromic Repeats - physiology
CRISPR
Electroporation
Gene-targeted knockout
Genetic Engineering
Genome
Molecular Sequence Data
PAX7
PAX7 Transcription Factor - physiology
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Summary: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. •We combined CRISPR and electroporation in the chicken embryo.•This led to efficient somatic genetic mutations in its tissues.•This completes the genetic toolbox to study development in amniotes.
ISSN:0012-1606
1095-564X
DOI:10.1016/j.ydbio.2015.08.007