基因敲除技术现状及应用

Status Quo and Application of Gene Knockout

功能基因组学的研究进展使基因敲除技术显得尤为重要。基因敲除技术从载体构建到细胞的筛选到动物模型的建立各方面都得到了发展。其中Cre-LoxP系统可以有效的控制打靶的发育阶段和组织类型,实现特定基因在特定时间和(或)空间的功能研究;转座子系统易于操作,所需时间短,具有高通量的特点,可以携带多种抗性标记,方便了同时进行多基因功能研究;基因捕获技术提供了获得基因敲除小鼠的高效方法,方便了进行小鼠基因组文库的研究。此外,进退策略、双置换法、标记和交换法、重组酶介导的盒子交换法也从不同方向发展了基因敲除技术。

Gene knockout is important for functional genomics. Great progress has been made in the vector construction of gene knockout, screening of aimed cells and animal models and the developments in these fields help to solve the problems in the study of genomic functions. Cre-LoxP system can effectively control the development stage and histology of gene knockout, thereby making it possible to study gene function in a given time and/or space. Transposon system is easy to manipulate, quick and can achieve high throughput, carry multiple resistance marker gene, which makes simuhaneous study of multiple genes possible. Gene knockout mice and can facilitate the research trapping provides a highly efficient method of obtain of mice genomic library. In addition, the techniques such as ＂hit and run＂, ＂double replacement＂, ＂tag and exchange＂ and ＂recombinase-mediated cassette exchange＂ all contribute to the development of gene knockout technology.