Due to sexual dimorphism in the growth of certain cultured fish species,the production of monosex fishes is desirable for the aquaculture industry.Nowadays,the most widely practiced technique available for the mass production of monosex fish populations is sex steroid-induced sex reversal.Here,a novel strategy for the successful production of all-female(AF)common carp(Cyprinus carpio L.),to take advan-tage of the sexual dimorphism in growth documented in this species,has been developed using genetic engineering via single gene-targeting manipulation without any exogenous hormone treatments.Male and female heterozygous cyp 17a1-deficient common carp were first obtained using the clustered regu-larly interspaced short palindromic repeats/CRISPR-associated endonuclease 9(CRISPR/Cas9)technique.An all-male phenotype for homozygous cyp 17a1-deficient carp,regardless of the individuals'sex-determination genotypes(XY or XX),has been observed.A male-specific DNA marker newly identified in our laboratory was used to screen the neomale carp population with the XX genotype from the cyp 17a 1-deficient carp.These neomale carp develop a normal testis structure with normal spermatoge-nesis and sperm capacity.The neomale common carp were then mated with wild-type(WT)females(cyp17a1+/+XX genotype)using artificial fertilization.All the AF offspring sample fish from the neomale-WT female mating were confirmed as having the cyp17a1+/-XX genotype,and normal develop-ment of gonads to ovaries was observed in 100.00％of this group at eight months post-fertilization(mpf).A total of 1000 carp fingerlings,500 from the WT male and female and 500 from the neomale and WT female mating,were mixed and reared in the same pond.The average body weight of cyp17a1+/-XX females was higher by 6.60％(8 mpf)and 32.66％(12 mpf)than that of the control common carp.Our study demonstrates the first successful production of a monosex teleost population with the advantages of sexual dimorphism in growth using genetic manipulation targeting a single locus.

Gang Zhai;Tingting Shu;Kuangxin Chen;Qiyong Lou;Jingyi Jia;Jianfei Huang;Chuang Shi;Xia Jin;Jiangyan He;Donghuo Jiang;Xueqiao Qian;Wei Hu;Zhan Yin

State Key Laboratory of Freshwater Ecology and Biotechnology,Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan 430072,China;College of Advanced Agricultural Sciences,University of Chinese Academy of Sciences,Beijing 100101,China;College of Fisheries,Huazhong Agricultural University,Wuhan 430070,China;HAID Research Institute,Guangdong HAID Group Co.,Ltd.,Guangzhou 511400,China;The Innovative Academy for Seed Design,Chinese Academy of Sciences,Beijing 100101,China

工程（英文）

[1]Gang Zhai;Tingting Shu;Kuangxin Chen;Qiyong Lou;Jingyi Jia;Jianfei Huang;Chuang Shi;Xia Jin;Jiangyan He;Donghuo Jiang;Xueqiao Qian;Wei Hu;Zhan Yin-.Successful Production of an All-Female Common Carp(Cyprinus carpio L.)Population Using cyp17a1-Deficient Neomale Carp)[J].工程（英文）,2022(01):181-189

Carp,cyp17a1,Neomale,monosex,cyp,17a1,neomale,spermatoge,cyp17a1+,+XX,mpf,fingerlings

Successful,Production,All,Female,Common,Cyprinus,carpio,Population,Using,Deficient,Due,sexual,dimorphism,growth,certain,cultured,species,production,fishes,desirable,aquaculture,industry,Nowadays,most,widely,practiced,technique,available,mass,populations,steroid,induced,reversal,Here,novel,strategy,successful,all,AF,common,take,documented,this,has,been,developed,using,genetic,engineering,via,single,targeting,manipulation,without,any,exogenous,hormone,treatments,Male,heterozygous,deficient,were,first,obtained,clustered,regu,larly,interspaced,short,palindromic,repeats,CRISPR,associated,endonuclease,Cas9,An,phenotype,homozygous,regardless,individuals,determination,genotypes,XY,observed,specific,marker,newly,identified,our,laboratory,was,used,screen,from,These,normal,testis,structure,nesis,capacity,then,mated,wild,WT,females,artificial,fertilization,offspring,sample,mating,confirmed,having,gonads,ovaries,group,months,post,total,mixed,reared,same,pond,average,body,weight,higher,by,than,that,control,Our,study,demonstrates,teleost,advantages,locus

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