title: Efficient dual sgRNA-directed large gene deletion in rabbit with CRISPR/Cas9 system.
abstract: The CRISPR RNA-guided Cas9 nuclease gene-targeting system has been extensively used to edit the genome of several organisms. However, most mutations reported to date have been are indels, resulting in multiple mutations and numerous alleles in targeted genes. In the present study, a large deletion of 105 kb in the TYR (tyrosinase) gene was generated in rabbit via a dual sgRNA-directed CRISPR/Cas9 system. The typical symptoms of albinism accompanied significantly decreased expression of TYR in the TYR knockout rabbits. Furthermore, the same genotype and albinism phenotype were found in the F1 generation, suggesting that large-fragment deletions can be efficiently transmitted to the germline and stably inherited in offspring. Taken together, our data demonstrate that mono and biallelic large deletions can be achieved using the dual sgRNA-directed CRISPR/Cas9 system. This system produces no mosaic mutations or off-target effects, making it an efficient tool for large-fragment deletions in rabbit and other organisms.
authors.affiliation: Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, 130062, China.
authors.forename: Yuning
authors.initials: Y
authors.lastname: Song
authors.affiliation: Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, 130062, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Yuan
authors.affiliation: Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, 130062, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, 130062, China.
authors.forename: Mao
authors.initials: M
authors.lastname: Chen
authors.affiliation: Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, 130062, China.
authors.forename: Jichao
authors.initials: J
authors.lastname: Deng
authors.affiliation: Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, 130062, China.
authors.forename: Qingyan
authors.initials: Q
authors.lastname: Lv
authors.affiliation: Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, 130062, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Sui
authors.affiliation: Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, 130062, China. lizj_1998@jlu.edu.cn.
authors.forename: Zhanjun
authors.initials: Z
authors.lastname: Li
authors.affiliation: Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, 130062, China. lai_liangxue@gibh.ac.cn.
authors.forename: Liangxue
authors.initials: L
authors.lastname: Lai
mesh_terms: D000417:Albinism / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D064113:CRISPR-Cas Systems; D005260:Female; D017353:Gene Deletion; D055786:Gene Knockout Techniques / Q000379:methods*; D014442:Monophenol Monooxygenase / Q000235:genetics*; D010641:Phenotype; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics; D011817:Rabbits / Q000196:embryology / Q000235:genetics*; D015053:Zygote / Q000378:metabolism
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems; D014442:Monophenol Monooxygenase
keywords: Albinism; CRISPR/Cas9; Rabbit; Tyrosinase
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