| 胞质内精子注射法介导外源DNA生产转基因动物 |
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| 引用本文： | 王荣祥,刘国辉,徐旭俊,陈建泉,成国祥.胞质内精子注射法介导外源DNA生产转基因动物[J].上海交通大学学报(农业科学版),2012,30(2):60-65. |
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| 作者姓名： | 王荣祥 刘国辉 徐旭俊 陈建泉 成国祥 |
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| 作者单位： | 上海转基因研究中心;上海杰隆生物工程股份有限公司,上海201210 |
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| 基金项目： | 转基因生物新品种培育科技重大专项(2009ZX08008-011B,2009ZX08011-032B);上海转基因动物育种与制药工程技术研究中心(10DZ2251700) |
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| 摘 要： | 通过胞质内精子注射法介导转基因已经成为生产转基因动物的一种有效手段。然而传统的ICSI介导法用冻融精子或Triton X-100处理的精子作为携带外源DNA的载体,经胚胎移植后生产转基因后代的效率很低,主要原因为这些处理方式在提高转染效率的同时对精子染色体造成了较大程度的损伤。最近研究者们提出了一种使用高浓度碱液对精子进行简单的预处理,高效地生产转基因后代的方法。这些精子完全失去了质膜与顶体,但仍保留有核的完整性,并且可以高效的将外源基因插入宿主基因组,比传统的精子处理方法获得了更理想的转基因动物的生产效率。
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| 关 键 词： | 胞质内精子注射 精子介导的转基因 转基因效率 DNA损伤 |
Production of Transgenic Offspring by Intracytoplasmic Sperm Injection (ICSI)-Mediated Exogenous DNA Gene Transfer |
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| WANG Rong-xiang
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LIU Guo-hui
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XU Xu-jun
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CHEN Jian-quan
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CHENG Guo-xiang.Production of Transgenic Offspring by Intracytoplasmic Sperm Injection (ICSI)-Mediated Exogenous DNA Gene Transfer[J].Journal of Shanghai Jiaotong University (Agricultural Science),2012,30(2):60-65. |
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| Authors: | WANG Rong-xiang LIU Guo-hui XU Xu-jun CHEN Jian-quan CHENG Guo-xiang |
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| Institution: | (Shanghai Transgenic Research Center;Shanghai Genon Bioengineering Co.Ltd,Shanghai 201210,China) |
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| Abstract: | Intracytoplasmic sperm injection(ICSI)-mediated transgenesis has been an effective measure for generating transgenic animals.However,the original ICSI-mediated methods that introduce exogenous gene using freeze-thawed or Triton X-100 treated spermatozoa as vector showed low offspring rates after embryo transfer.Severe paternal chromosomal damage is the primary cause of decreasing in embryo development whereas the transfection efficiency increased using these treatments.Recently,researchers introduced an improved method for generating transgenic pups efficiently using a simple pretreatment of spermatozoa with highly alkaline medium.These spermatozoa lost their plasma membrane and acrosome completely,while still maintained nuclear integrity,which results in an efficient insertion of an exogenous gene into the host genome.The improved method is more efficient in generating transgenic offspring than classical ones. |
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| Keywords: | intracytoplasmic sperm injection(ICSI) sperm mediated gene transfer(SMGT) transgenic efficiency DNA damage |
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