基因组靶向修饰技术研究进展

摘要

Targeted genome modification technology is one of the important methods to study genes′function, which also can be used for the treatment of human diseases at the same time.It is becoming a hot field of biology research in recent years.Because of its low efficiency and toxicity, traditional targeted modification technology will be replaced by a more efficient and safer technology.Thus three generations of targeted genome modification technologies have been developed.It includes ZFN, TALEN and CRISPR/Cas9.These three techniques overcome the defects of traditional technology.Here, we discuss the research progress in the structure, mechanism and application of these three technologies, especially newly developed CRISPR/Cas9.Finally, the three generations of genome modifi-cation techniques are compared.%基因组靶向修饰技术是研究基因功能的重要方法之一，该技术也被用于人类疾病的治疗上，从而成为近来生物学研究的热点。传统的靶向修饰技术由于其效率低、有毒性等缺点注定其将要被更高效、安全的技术所取代，因此产生了后来的三代基因组靶向修饰技术：锌指核酸酶（ Zinc finger nuclease， ZFN）、类转录激活因子效应物核酸酶（ Transcription activator－like effector nuclease， TALEN）和常间回文重复序列丛集关联蛋白系统（ Clustered regularly interspaced short palindromic repeats／CRISPR－associated proteins 9， CRISPR／Cas9）。这3种技术在克服传统技术缺陷的基础上，也针对其上一代技术的缺陷进行了自身的改善。对三代基因组靶向修饰技术，尤其最近发展起来的CRISPR／Cas9的结构组成、作用原理和基因定点修饰中的应用进行阐述，最后对三代基因组靶向修饰技术进行比较。