State Key Laboratory of Brain and Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Datun Road 15, Beijing 100101, China.
J Genet Genomics. 2013 Jun 20;40(6):281-9. doi: 10.1016/j.jgg.2013.03.013. Epub 2013 Apr 12.
Precise modifications of complex genomes at the single nucleotide level have been one of the big goals for scientists working in basic and applied genetics, including biotechnology, drug development, gene therapy and synthetic biology. However, the relevant techniques for making these manipulations in model organisms and human cells have been lagging behind the rapid high throughput studies in the post-genomic era with a bottleneck of low efficiency, time consuming and laborious manipulation, and off-targeting problems. Recent discoveries of TALEs (transcription activator-like effectors) coding system and CRISPR (clusters of regularly interspaced short palindromic repeats) immune system in bacteria have enabled the development of customized TALENs (transcription activator-like effector nucleases) and CRISPR/Cas9 to rapidly edit genomic DNA in a variety of cell types, including human cells, and different model organisms at a very high efficiency and specificity. In this review, we first briefly summarize the development and applications of TALENs and CRISPR/Cas9-mediated genome editing technologies; compare the advantages and constraints of each method; particularly, discuss the expected applications of both techniques in the field of site-specific genome modification and stem cell based gene therapy; finally, propose the future directions and perspectives for readers to make the choices.
精确修饰复杂基因组的单个核苷酸水平一直是从事基础和应用遗传学研究的科学家(包括生物技术、药物开发、基因治疗和合成生物学)的主要目标之一。然而,在基因组后时代,相关技术在模型生物和人类细胞中的应用一直滞后于高通量快速研究,其效率低、耗时费力、靶向问题严重。最近在细菌中发现了 TALEs(转录激活样效应物)编码系统和 CRISPR(规律成簇间隔短回文重复)免疫系统,这使得定制 TALENs(转录激活样效应物核酸酶)和 CRISPR/Cas9 的开发成为可能,从而能够在各种细胞类型(包括人类细胞和不同的模型生物)中以非常高的效率和特异性快速编辑基因组 DNA。在这篇综述中,我们首先简要总结了 TALENs 和 CRISPR/Cas9 介导的基因组编辑技术的发展和应用;比较了每种方法的优缺点;特别讨论了这两种技术在特定基因组修饰和基于干细胞的基因治疗领域的预期应用;最后,为读者提供了未来的方向和观点,以便做出选择。