Successful production of an all-female common carp (Cyprinus carpio L.) population using cyp17a1-deficient neomale carp
Successful production of an all-female common carp (Cyprinus carpio L.) population using cyp17a1-deficient neomale carp
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DOI：
10.1016/j.eng.2021.03.026
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发表时间：
2021-08
期刊：
影响因子：
12.8
通讯作者：
Gang Zhai;Tingting Shu;Kuangxin Chen;Qi-yong Lou;Jingyi Jia;Jianfei Huang;Chuang Shi;Xia Jin
中科院分区：
文献类型：
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作者：
Gang Zhai;Tingting Shu;Kuangxin Chen;Qi-yong Lou;Jingyi Jia;Jianfei Huang;Chuang Shi;Xia Jin
由于某些养殖鱼类在生长过程中的两性异形，单性别鱼类的生产对于水产养殖业是理想的。目前，大规模生产单性鱼类种群最广泛采用的技术是性类固醇诱导的性逆转。在这里，一种新的策略，成功地生产全雌性（AF）普通鲤鱼（Cyprinus carpioL.），为了利用在该物种中记录的生长中的两性异形，已经使用基因工程通过单基因靶向操作而不使用任何外源激素处理来开发。首先使用成簇规则间隔短回文重复序列/CRISPR相关内切核酸酶9（CRISPR/Cas9）技术获得雄性和雌性杂合酶17 a1缺陷型鲤鱼。无论个体的性别决定基因型（XY或XX），均观察到纯合yp 17 a1缺陷鲤鱼的全雄性表型。利用本实验室新发现的一个雄性特异DNA标记，从yp 17 a1基因缺失的新鲤群体中筛选出XX基因型的新鲤群体。这些新雄性鲤鱼发育正常的睾丸结构，具有正常的精子发生和精子能力。采用人工受精的方法将新鲤与野生型（WT）鲤（cyp 17 a1 +/+XX基因型）进行交配。所有来自新雄性-WT雌性交配的AF后代样品鱼被确认为具有yp 17 a1 +/−XX基因型，并且在受精后8个月（mpf）时，在该组中观察到100.00%的性腺至卵巢的正常发育。共1000尾鲤鱼鱼种，500尾来自WT雄性和雌性，500尾来自新雄性和WT雌性交配，在同一池塘中混合饲养。cyp 17 a1 +/-XX雌鲤的平均体重比对照鲤高6.60%（8 mpf）和32.66%（12 mpf）。我们的研究表明，第一次成功地生产的单性硬骨鱼种群的优势，性二型的增长，利用遗传操作针对一个单一的位点。
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 carpioL.), to take advantage 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 heterozygouscyp17a1-deficient common carp were first obtained using the clustered regularly interspaced short palindromic repeats/CRISPR-associated endonuclease 9 (CRISPR/Cas9) technique. An all-male phenotype for homozygouscyp17a1-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 thecyp17a1-deficient carp. These neomale carp develop a normal testis structure with normal spermatogenesis 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 thecyp17a1+/−XX genotype, and normal development 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 ofcyp17a1+/−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.