Germline Gene Editing in Chickens by Efficient CRISPR-Mediated Homologous Recombination in Primordial Germ Cells

被引:77
作者
Dimitrov, Lazar [1 ]
Pedersen, Darlene [1 ]
Ching, Kathryn H. [1 ]
Yi, Henry [1 ]
Collarini, Ellen J. [1 ]
Izquierdo, Shelley [1 ]
van de lavoir, Marie-Cecile [1 ]
Leighton, Philip A. [1 ]
机构
[1] Crystal Biosci Inc, Emeryville, CA USA
关键词
KNOCKOUT; SYSTEM; BIRDS;
D O I
10.1371/journal.pone.0154303
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The CRISPR/Cas9 system has been applied in a large number of animal and plant species for genome editing. In chickens, CRISPR has been used to knockout genes in somatic tissues, but no CRISPR-mediated germline modification has yet been reported. Here we use CRISPR to target the chicken immunoglobulin heavy chain locus in primordial germ cells (PGCs) to produce transgenic progeny. Guide RNAs were co-transfected with a donor vector for homology-directed repair of the double-strand break, and clonal populations were selected. All of the resulting drug-resistant clones contained the correct targeting event. The targeted cells gave rise to healthy progeny containing the CRISPR-targeted locus. The results show that gene-edited chickens can be obtained by modifying PGCs in vitro with the CRISPR/Cas9 system, opening up many potential applications for efficient genetic modification in birds.
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页数:10
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