White Martyn K, Kaminski Rafal, Young Won-Bin, Roehm Pamela C, Khalili Kamel
Center for Neurovirology and Comprehensive NeuroAIDS Center, Department of Neuroscience, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, Philadelphia, Pennsylvania, 19140.
J Cell Biochem. 2017 Nov;118(11):3586-3594. doi: 10.1002/jcb.26099. Epub 2017 Jul 4.
The CRISPR or clustered regularly interspaced short palindromic repeats system is currently the most advanced approach to genome editing and is notable for providing an unprecedented degree of specificity, effectiveness, and versatility in genetic manipulation. CRISPR evolved as a prokaryotic immune system to provide an acquired immunity and resistance to foreign genetic elements such as bacteriophages. It has recently been developed into a tool for the specific targeting of nucleotide sequences within complex eukaryotic genomes for the purpose of genetic manipulation. The power of CRISPR lies in its simplicity and ease of use, its flexibility to be targeted to any given nucleotide sequence by the choice of an easily synthesized guide RNA, and its ready ability to continue to undergo technical improvements. Applications for CRISPR are numerous including creation of novel transgenic cell animals for research, high-throughput screening of gene function, potential clinical gene therapy, and nongene-editing approaches such as modulating gene activity and fluorescent tagging. In this prospect article, we will describe the salient features of the CRISPR system with an emphasis on important drawbacks and considerations with respect to eliminating off-target events and obtaining efficient CRISPR delivery. We will discuss recent technical developments to the system and we will illustrate some of the most recent applications with an emphasis on approaches to eliminate human viruses including HIV-1, JCV and HSV-1 and prospects for the future. J. Cell. Biochem. 118: 3586-3594, 2017. © 2017 Wiley Periodicals, Inc.
CRISPR(成簇规律间隔短回文重复序列)系统是目前基因组编辑领域最先进的方法,其在基因操作中具有前所未有的特异性、有效性和多功能性,因而备受瞩目。CRISPR最初作为原核生物的免疫系统而进化,用于提供获得性免疫,抵抗诸如噬菌体等外来遗传元件。最近,它已发展成为一种用于特异性靶向复杂真核基因组中核苷酸序列以进行基因操作的工具。CRISPR的强大之处在于其简单易用,通过选择易于合成的向导RNA可灵活靶向任何给定的核苷酸序列,并且能够持续进行技术改进。CRISPR的应用广泛,包括创建用于研究的新型转基因细胞动物、高通量筛选基因功能、潜在的临床基因治疗以及诸如调节基因活性和荧光标记等非基因编辑方法。在这篇前瞻性文章中,我们将描述CRISPR系统的显著特征,重点关注在消除脱靶事件和实现高效CRISPR递送方面的重要缺点及注意事项。我们将讨论该系统最近的技术发展,并举例说明一些最新应用,重点是消除包括HIV-1、JCV和HSV-1在内的人类病毒的方法以及未来前景。《细胞生物化学杂志》118: 3586 - 3594, 2017。© 2017威利期刊公司