摘要
通过对基因组特定区域进行精确定向遗传修饰,一方面可以针对目标序列进行精确突变,获得突变材料,对目标基因功能进行明确鉴定;另一方面可以进行目标序列的精确置换或插入,将外源基因随机导入造成的表达及遗传的不确定性降至最低。传统的基因定向修饰技术仅依赖于细胞自身的同源重组,修饰效率低下,而且还存在位置效应和遗传不稳定等诸多问题。通过引入序列特异性核酸酶(sequence-specific nucleases,SSN),可以在基因组特定位点造成DNA双链断裂(double strand break,DSB),促进依赖于细胞内源"同源重组"及"非同源末端连接"的DNA修复事件的定向发生,实现基因组定向遗传修饰效率的大幅提升。迄今为止,在基因组定向遗传修饰研究及应用领域,已经有多种不同类型的序列特异性核酸酶被有效使用,在多种生物中实现了不同类型的基因组定向遗传修饰。该文首先综述了SSN的结构特征及技术原理,然后对SSN技术在植物基因组定向遗传修饰中的研究进展和应用前景进行了重点介绍。
Through gene targeting at specific loci of the genome, we can obtain mutant which have precise mutation at target sequence and clearly identify the gene function. On the other hand, genome targeting by directly substitution or insertion, can minimize the uncertainty caused by the expression of foreign gene which inserted randomly. The traditional gene targeting technique, which rely solely on the cell＇s own homologous recombination, is inefficient. And there also are many other issues, like position effects and genetic instability, and so on. By introducing the sequence-specific nucleases （SSN）, we can introduce DNA double strand break at specific genomic loci, stimulate the occurrence of the direct DNA repair events which depend on intracellular ＂homologous recombination＂ and ＂non-homologous end joining＂, and dramatically improve the efficiency of gene targeting finally. To date, at the area of genome targeting research, there are different types of sequence-specific nucleases that have been effectively used. And in multiple species different types of target genetic modifications have been achieved. In this review, the principles and structural characteristics of SSN were introduced firstly, and then we discussed the advances and prospects of genome targeting in plants
出处
《中国细胞生物学学报》
CAS
CSCD
北大核心
2013年第11期1650-1659,共10页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:30900779
31271420)
国家重点基础研究发展计划(973)项目(批准号:2009CB119105)
天津市自然科学基金(批准号:13JCZDJC29000)资助的课题~~
