Qin Guyu, Qin Zhenkui, Lu Cuiyu, Ye Zhi, Elaswad Ahmed, Jin Yulin, Khan Mohd Golam Quader, Su Baofeng, Dunham Rex A
School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL 36849, USA.
Biology (Basel). 2023 Mar 1;12(3):392. doi: 10.3390/biology12030392.
Follicle-stimulating hormone () plays an important role in sexual maturation in catfish. Knocking out the gene in the fish zygote should suppress the reproduction of channel catfish (). In this study, transcription activator-like effector nuclease (TALEN) plasmids targeting the gene were electroporated into fertilized eggs with the standard double electroporation technique. Targeted cleavage efficiency was 63.2% in P-knockout catfish. Ten of fifteen (66.7%) control pairs spawned, and their eggs had 32.3-74.3% average hatch rates in 2016 and 2017. Without hormone therapy, the spawning rates of P mutants ranged from 33.3 to 40.0%, with an average egg hatching rate of 0.75%. After confirmation of the low fertility of P mutants in 2016, human chorionic gonadotropin (HCG) hormone therapy improved the spawning rates by 80% for female mutants and 88.9% for male mutants, and the mean hatch rate was 35.0% for F embryos, similar to that of the controls ( > 0.05). Polymerase chain reaction (PCR) identification showed no potential TALEN plasmid integration into the P channel catfish genome. Neither the P nor the F mutant fish showed any noticeable changes in in body weight, survival rate, and hatching rate when the reproductive gene was knocked out. F families had a mean inheritance rate of 50.3%. The results brought us one step closer to allowing implementation of certain genetic techniques to aquaculture and fisheries management, while essentially eliminating the potential environment risk posed by transgenic, hybrid, and exotic fish as well as domestic fish.
促卵泡激素（FSH）在鲶鱼的性成熟过程中起着重要作用。敲除鱼受精卵中的FSH基因应该会抑制斑点叉尾鮰的繁殖。在本研究中，使用标准的双电穿孔技术将靶向FSH基因的转录激活样效应核酸酶（TALEN）质粒电穿孔导入受精卵中。在FSH基因敲除的鲶鱼中，靶向FSH的切割效率为63.2%。2016年和2017年，15对对照组中有10对产卵，其卵的平均孵化率为32.3%-74.3%。在没有激素治疗的情况下，FSH突变体的产卵率在33.3%至40.0%之间，平均卵孵化率为0.75%。在确认2016年FSH突变体的繁殖力较低后，人绒毛膜促性腺激素（HCG）激素治疗使雌性突变体的产卵率提高了80%，雄性突变体提高了88.9%，F1胚胎的平均孵化率为35.0%，与对照组相似（P>0.05）。聚合酶链反应（PCR）鉴定表明，没有TALEN质粒整合到斑点叉尾鮰基因组中。当生殖基因被敲除时，FSH突变体和F1突变体鱼在体重、存活率和孵化率方面均未表现出任何明显变化。F1家系的平均遗传率为50.3%。这些结果使我们更接近在水产养殖和渔业管理中应用某些遗传技术，同时基本消除转基因鱼、杂交鱼、外来鱼以及家养鱼类带来的潜在环境风险。
