Journal article
CRISPR/Cas9 Targets Chicken Embryonic Somatic Cells in Vitro and in Vivo and generates Phenotypic Abnormalities
KD Abu-Bonsrah, D Zhang, DF Newgreen
Scientific Reports | NATURE PORTFOLIO | Published : 2016
DOI: 10.1038/srep34524
Abstract
Chickens are an invaluable model for studying human diseases, physiology and especially development, but have lagged in genetic applications. With the advent of Programmable Engineered Nucleases, genetic manipulation has become efficient, specific and rapid. Here, we show that the CRISPR/Cas9 system can precisely edit the chicken genome. We generated HIRA, TYRP1, DICER, MBD3, EZH2, and 6 other gene knockouts in two chicken cell lines using the CRISPR/Cas9 system, with no off-target effects detected. We also showed that very large deletions (>75 kb) could be achieved. We also achieved targeted modification by homology-directed repair (HDR), producing MEN2A and MEN2B mutations of the RET gene...
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Awarded by National Institutes of Health
Funding Acknowledgements
We thank Damien Hudson (MCRI) and Craig Smith (MCRI) for the DT40 and DF-1 cells respectively, Marguerite Evans-Galea (MCRI) for the mouse brain and Yoshiko Takahashi (Kyoto, Japan) for Tol2 and transposase plasmids. We thank Matthew Burton and Paul Lau (Flow Cytometry-MCRI), Paul Kalitsis (Chromosomal Research-MCRI), Hwee Ong (Genetic Health Research-MCRI) and Alex Combes (Kidney Development, Disease and Regeneration) for their technical advice and support. The mCherry monoclonal antibody (3A11) was developed and supplied by the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242. This research was supported by NHMRC grant 1069757 and MCRI. HSCR-related constructs were contributed to by Foundation for Children 2014-211. We acknowledge the Victorian Government's Operational Infrastructure Support Program to MCRI. K.D.A.-B. holds an IPRS and APA(Int) PhD Scholarship through the Department of Paediatrics, University of Melbourne.