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 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.
EMBL Australia, Australian Regenerative Medicine Institute, Monash University, Building 75, Clayton, VIC 3800, Australia.
Nadège
N
Véron
EMBL Australia, Australian Regenerative Medicine Institute, Monash University, Building 75, Clayton, VIC 3800, Australia.
Zhengdong
Z
Qu
EMBL Australia, Australian Regenerative Medicine Institute, Monash University, Building 75, Clayton, VIC 3800, Australia.
Phoebe A S
PA
Kipen
EMBL Australia, Australian Regenerative Medicine Institute, Monash University, Building 75, Clayton, VIC 3800, Australia.
Claire E
CE
Hirst
EMBL Australia, Australian Regenerative Medicine Institute, Monash University, Building 75, Clayton, VIC 3800, Australia. Electronic address: christophe.marcelle@monash.edu.
Christophe
C
Marcelle
D000818:Animals; D001483:Base Sequence; D002642:Chick Embryo; D002645:Chickens; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000502:physiology*; D018274:Electroporation; D005818:Genetic Engineering; D016678:Genome; D008969:Molecular Sequence Data; D051763:PAX7 Transcription Factor / Q000502:physiology
D051763:PAX7 Transcription Factor
CRISPR; Cas9; Chicken; Electroporation; Gene-targeted knockout; PAX7