中国农业科学 ›› 2014, Vol. 47 ›› Issue (21): 4211-4223.doi: 10.3864/j.issn.0578-1752.2014.21.007
• 高效、安全、规模化转基因技术:机会与挑战 • 上一篇 下一篇
牟玉莲,阮进学,吴添文,程英,魏景亮,樊俊华,李奎
收稿日期:
2014-04-02
修回日期:
2014-06-27
出版日期:
2014-11-01
发布日期:
2014-11-01
通讯作者:
李奎,E-mail:likui@caas.cn
作者简介:
牟玉莲,E-mail:muyulian76@iascaas.net.cn
基金资助:
MU Yu-lian, RUAN Jin-xue, WU Tian-wen, CHENG Ying, WEI Jing-liang, FAN Jun-hua, LI Kui
Received:
2014-04-02
Revised:
2014-06-27
Online:
2014-11-01
Published:
2014-11-01
摘要: 猪是重要的经济动物,优良猪种的培育对农业的发展具有重要的作用。同时,作为一种常用的试验动物,猪在解剖学、生理学、遗传学等方面与人类高度相似,其在生命科学研究领域中也具有独特的应用价值。转基因技术可以运用基因工程等实验技术手段,对动物基因组进行有目的的遗传修饰,使修饰改造的基因稳定遗传给后代,从而获得满足人类特定需求的动物个体。因此,猪规模化转基因技术体系的建立对转基因育种、基因功能研究和人类疾病模型研究等方面具有重要的意义。本文回顾了国内外猪转基因技术体系的研究进展,总结了目标基因的选择原则,介绍了原核显微注射、体细胞核移植、锌指核酸酶(Zinc-finger nucleases, ZFN)、转录激活子样效应子核酸酶(Transcription activator-like effector nucleases, TALEN)和规律成簇间隔短回文重复序列/ Cas9(Clustered regularly interspaced short palindromic repeat, CRISPR/Cas9)系统、精子载体和逆转录病毒等各种目标基因导入手段,并分析其优缺点和适用范围,列举了表达调控策略中常用的基因调控元件和相关调控系统。近年来,随着转基因重大专项工作的推进,中国猪规模化转基因技术研究取得长足的发展:发掘出大批功能基因;搭建了ZFN、TALEN、CRISPR/Cas9介导的高效基因修饰技术平台及其切割效率检测平台;对供体细胞的分离、培养和筛选流程进行优化;完善体细胞核移植和原核显微注射体系平台,并与新型的基因组编辑技术有机结合,实现目的基因定点、高效、安全的整合;通过将再克隆、胚胎移植、人工受精及常规育种等多种技术的集成创新,建立转基因猪扩繁技术体系,可在较短时间内扩大转基因猪数量;并且实现了这些技术体系的资源共享,为众多单位提供了技术服务支撑,培训技术人员,使转基因技术得到普遍的应用。但是中国猪规模化转基因技术体系仍然存在着一系列的问题:决定转基因猪相关性状并具有自主知识产权的主效基因少;标记基因存在潜在的危害;外源基因随机整合率低、稳定性差;基因打靶等技术环节还不成熟;仔猪成活率低等仍然是猪规模化转基因技术发展面临的瓶颈。因此,要实现猪规模化转基因体系构建的技术性突破,必须结合相关组学工具,深入挖掘并获得具有自主知识产权的重要功能基因和调控元件;构建高效、安全的转多基因载体,实现外源基因的协同、高效表达;完善转基因猪制备过程中的相关技术环节。总之,在猪规模化转基因技术体系的支撑下,中国的转基因猪培育研究必将进入一个新的高速发展时期。
牟玉莲,阮进学,吴添文,程英,魏景亮,樊俊华,李奎. 猪规模化转基因技术体系构建及其应用[J]. 中国农业科学, 2014, 47(21): 4211-4223.
MU Yu-lian, RUAN Jin-xue, WU Tian-wen, CHENG Ying, WEI Jing-liang, FAN Jun-hua, LI Kui. Construction and Application of a Scale Transgenic Technology System for Pigs[J]. Scientia Agricultura Sinica, 2014, 47(21): 4211-4223.
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