【摘 要】
：
转录激活因子样效应物核酸酶(Transcription activator-like effector nucleases,TALEN)是目前最有发展前景的基因组修饰技术.Dmrt1是脊椎动物精巢分化和维持的关键基因.
【机 构】
：
西南大学生命科学学院,淡水鱼类资源与生殖发育教育部重点实验室,重庆,400175
【出 处】
：
2012年全国博生研究生学术论坛——可持续的水产养殖：理论与实践
论文部分内容阅读
转录激活因子样效应物核酸酶(Transcription activator-like effector nucleases,TALEN)是目前最有发展前景的基因组修饰技术.Dmrt1是脊椎动物精巢分化和维持的关键基因.本研究在养殖鱼类尼罗罗非鱼(Oreochromis niloticus)建立了TALEN靶向基因敲除技术.通过对Drnrt1 基因敲除结果发现,TALEN能高效(约37％)的对Dmrt1基因进行编辑,包括删除不同碱基数目(7、9、10等)的突变类型.4月龄Dmrt1敲除个体性腺组织学观察显示精巢不能正常发育,如不能正常起始减数分裂,因此少有甚至没有对照组精巢中所见的精母细胞、精细胞和精子,仅见大量精原细胞,部分退化区域甚至出现卵母细胞,同时,未见曲细精管.免疫组化结果显示Dmrt1敲除个体精巢中检测到芳香化酶(合成雌激素的关建酶)的表达,而没有检测到胆固醇侧链裂解酶P450scc的表达.以上结果表明像哺乳动物一样,Dmrt1在硬骨鱼类精巢的正常分化、发育以及精原细胞减数分裂起始中起重要作用.
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