title: Targeted disruption of sp7 and myostatin with CRISPR-Cas9 results in severe bone defects and more muscular cells in common carp.
abstract: The common carp (Cyprinus carpio) as one of the most important aquaculture fishes produces over 3 million metric tones annually, approximately 10% the annual production of the all farmed freshwater fish worldwide. However, the tetraploidy genome and long generation-time of the common carp have made its breeding and genetic studies extremely difficult. Here, TALEN and CRISPR-Cas9, two versatile genome-editing tools, are employed to target common carp bone-related genes sp7, runx2, bmp2a, spp1, opg, and muscle suppressor gene mstn. TALEN were shown to induce mutations in the target coding sites of sp7, runx2, spp1 and mstn. With CRISPR-Cas9, the two common carp sp7 genes, sp7a and sp7b, were mutated individually, all resulting in severe bone defects; while mstnba mutated fish have grown significantly more muscle cells. We also employed CRISPR-Cas9 to generate double mutant fish of sp7a;mstnba with high efficiencies in a single step. These results demonstrate that both TALEN and CRISPR-Cas9 are highly efficient tools for modifying the common carp genome, and open avenues for facilitating common carp genetic studies and breeding.
authors.affiliation: Center for Circadian Clocks, Soochow University, Suzhou 215123, Jiangsu, China.|School of Biology &Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, Jiangsu, China.
authors.forename: Zhaomin
authors.initials: Z
authors.lastname: Zhong
authors.affiliation: Center for Circadian Clocks, Soochow University, Suzhou 215123, Jiangsu, China.|School of Biology &Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, Jiangsu, China.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Niu
authors.affiliation: Center for Circadian Clocks, Soochow University, Suzhou 215123, Jiangsu, China.|School of Biology &Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, Jiangsu, China.
authors.forename: Mingyong
authors.initials: M
authors.lastname: Wang
authors.affiliation: Center for Circadian Clocks, Soochow University, Suzhou 215123, Jiangsu, China.|School of Biology &Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, Jiangsu, China.
authors.forename: Guodong
authors.initials: G
authors.lastname: Huang
authors.affiliation: Center for Circadian Clocks, Soochow University, Suzhou 215123, Jiangsu, China.|School of Biology &Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, Jiangsu, China.
authors.forename: Shuhao
authors.initials: S
authors.lastname: Xu
authors.affiliation: Center for Circadian Clocks, Soochow University, Suzhou 215123, Jiangsu, China.|School of Biology &Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, Jiangsu, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Center for Circadian Clocks, Soochow University, Suzhou 215123, Jiangsu, China.|School of Biology &Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, Jiangsu, China.
authors.forename: Xiaona
authors.initials: X
authors.lastname: Xu
authors.affiliation: School of Biology &Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, Jiangsu, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Hou
authors.affiliation: Heilongjiang River Fisheries Research Institute of Chinese Academy of Fishery Sciences, Harbin, China.
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Sun
authors.affiliation: Center for Circadian Clocks, Soochow University, Suzhou 215123, Jiangsu, China.|School of Biology &Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, Jiangsu, China.|Institute of Neuroscience, University of Oregon, Eugene, Oregon 97403, USA.
authors.forename: Yilin
authors.initials: Y
authors.lastname: Yan
authors.affiliation: Center for Circadian Clocks, Soochow University, Suzhou 215123, Jiangsu, China.|School of Biology &Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, Jiangsu, China.
authors.forename: Han
authors.initials: H
authors.lastname: Wang
mesh_terms: D000818:Animals; D001483:Base Sequence; D001842:Bone and Bones / Q000378:metabolism* / Q000473:pathology; D064113:CRISPR-Cas Systems; D002347:Carps / Q000235:genetics* / Q000378:metabolism; D029941:Fish Proteins / Q000235:genetics*; D000072669:Gene Editing / Q000379:methods; D008957:Models, Genetic; D009132:Muscles / Q000166:cytology / Q000378:metabolism*; D009154:Mutation; D055435:Myostatin / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D029941:Fish Proteins; D055435:Myostatin; D014157:Transcription Factors