基因修饰技术研究进展

Advances in genetic modification technologies

摘要:

基因修饰技术是用于基因组定点改造的分子工具，目前主要有锌指核酸酶 (ZFN) 技术、转录激活子样效应物核酸酶 (TALEN) 技术和CRISPR-Cas核酸酶 (CRISPR-Cas) 技术。这些核酸酶都可以在DNA靶位点产生双链断裂 (DSB)，诱发细胞内源性的修复机制，激活体内非同源末端连接 (NHEJ) 或同源重组 (HR) 两种不同的修复机制，从而实现内源基因的敲除或外源基因的定点敲入。近年来，基因修饰技术已成功应用到细菌、酵母、人类细胞、果蝇、斑马鱼、小鼠、大鼠、家畜、食蟹猴、拟南芥、水稻、烟草、玉米、高粱、小麦和大麦等多种生物，显示了其强大的基因编辑优势。特别是新近出现的CRISPR-Cas9技术，降低了成本，使基因编辑变得简洁、高效和易于操作，得到了很多研究人员的关注。本文系统介绍了以上3种技术的原理及最新研究进展，并对未来的研究和应用做出了展望。

Abstract:

Genetic modification technology is a new molecular tool for targeted genome modification. It includes zinc finger nucleases (ZFN) technology, transcription activator-like effector nucleases (TALEN) technology and clustered regularly interspaced short palindromic repeat (CRISPR)-associated (Cas) (CRISPR-Cas) nucleases technology. All of these nucleases create DNA double-strand breaks (DSB) at chromosomal targeted sites and induce cell endogenous mechanisms that are primarily repaired by the non-homologous end joining (NHEJ) or homologous recombination (HR) pathway, resulting in targeted endogenous gene knock-out or exogenous gene insertion. In recent years, genetic modification technologies have been successfully applied to bacteria, yeast, human cells, fruit fly, zebra fish, mouse, rat, livestock, cynomolgus monkey, Arabidopsis, rice, tobacco, maize, sorghum, wheat, barley and other organisms, showing its enormous advantage in gene editing field. Especially, the newly developed CRISPR-Cas9 system arose more attention because of its low cost, high effectiveness, simplicity and easiness. We reviewed the principles and the latest research progress of these three technologies, as well as prospect of future research and applications.