TALEN和CRISPR/Cas基因组编辑系统:发现工具
TALEN and CRISPR/Cas Genome Editing Systems: Tools of Discovery.
作者信息
Nemudryi A A, Valetdinova K R, Medvedev S P, Zakian S M
机构信息
Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Lavrentyev Prosp., 10, Novosibirsk, Russia, 630090 ; Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentyev Prosp., 8, Novosibirsk, Russia, 630090 ; Meshalkin Novosibirsk State Research Institute of Circulation Pathology, Ministry of Health of the Russian Federation, Rechkunovskaya Str., 15, Novosibirsk, Russia, 630055.
Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Lavrentyev Prosp., 10, Novosibirsk, Russia, 630090 ; Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentyev Prosp., 8, Novosibirsk, Russia, 630090 ; Meshalkin Novosibirsk State Research Institute of Circulation Pathology, Ministry of Health of the Russian Federation, Rechkunovskaya Str., 15, Novosibirsk, Russia, 630055 ; Novosibirsk State University, Pirogova Str., 2, Novosibirsk, Russia, 630090.
出版信息
Acta Naturae. 2014 Jul;6(3):19-40.
Precise studies of plant, animal and human genomes enable remarkable opportunities of obtained data application in biotechnology and medicine. However, knowing nucleotide sequences isn't enough for understanding of particular genomic elements functional relationship and their role in phenotype formation and disease pathogenesis. In post-genomic era methods allowing genomic DNA sequences manipulation, visualization and regulation of gene expression are rapidly evolving. Though, there are few methods, that meet high standards of efficiency, safety and accessibility for a wide range of researchers. In 2011 and 2013 novel methods of genome editing appeared - this are TALEN (Transcription Activator-Like Effector Nucleases) and CRISPR (Clustered Regulatory Interspaced Short Palindromic Repeats)/Cas9 systems. Although TALEN and CRISPR/Cas9 appeared recently, these systems have proved to be effective and reliable tools for genome engineering. Here we generally review application of these systems for genome editing in conventional model objects of current biology, functional genome screening, cell-based human hereditary disease modeling, epigenome studies and visualization of cellular processes. Additionally, we review general strategies for designing TALEN and CRISPR/Cas9 and analyzing their activity. We also discuss some obstacles researcher can face using these genome editing tools.
对植物、动物和人类基因组进行精确研究,能为生物技术和医学领域提供大量利用所获数据的机会。然而,仅了解核苷酸序列不足以理解特定基因组元件的功能关系及其在表型形成和疾病发病机制中的作用。在后基因组时代,能够对基因组DNA序列进行操作、可视化以及调控基因表达的方法正在迅速发展。不过,能满足广泛研究人员对效率、安全性和可及性高标准要求的方法却很少。2011年和2013年出现了新型基因组编辑方法——即转录激活样效应因子核酸酶(TALEN)和规律成簇间隔短回文重复序列(CRISPR)/Cas9系统。尽管TALEN和CRISPR/Cas9出现时间不长,但已证明它们是基因组工程的有效且可靠工具。在此,我们全面综述这些系统在当前生物学常规模式生物中的基因组编辑、功能基因组筛选、基于细胞的人类遗传疾病建模、表观基因组研究以及细胞过程可视化方面的应用。此外,我们还综述了设计TALEN和CRISPR/Cas9以及分析其活性的一般策略。我们也讨论了研究人员在使用这些基因组编辑工具时可能面临的一些障碍。