Choudhary Eira, Lunge Ajitesh, Agarwal Nisheeth
Translational Health Science and Technology Institute, NCR Biotech Science Cluster, 3rd Milestone, Faridabad-Gurgaon Expressway, Faridabad 121001, Haryana, India; Symbiosis School of Biomedical Sciences, Symbiosis International University, Lavale, Pune 412115, Maharashtra, India.
Translational Health Science and Technology Institute, NCR Biotech Science Cluster, 3rd Milestone, Faridabad-Gurgaon Expressway, Faridabad 121001, Haryana, India; Jawaharlal Nehru University, New Mehrauli Road, Near Munirka, New Delhi 110067, Delhi, India.
Tuberculosis (Edinb). 2016 May;98:132-8. doi: 10.1016/j.tube.2016.03.005. Epub 2016 Mar 26.
Tremendous amount of physiological and functional complexities acquired through decades of evolutionary pressure makes Mycobacterium tuberculosis (Mtb) one of the most dreadful microorganisms infecting humans from centuries. Astonishing advances in genomics and genome editing tools substantially grew our knowledge about Mtb as an organism but dramatically failed to completely understand it as a pathogen. Though conventional tools based on homologous recombination, antisense, controlled proteolysis, etc. have made important contributions in advancing our understanding of the pathophysiology of Mtb, yet these approaches have not accentuated our exploration of mycobacterium on account of certain technical limitations. In this review article we have compiled various approaches implemented in genome editing of mycobacteria along with the latest adaptation of clustered regularly interspaced short palindromic repeat (CRISPR)-interference (CRISPRi), emphasizing the achievements and challenges associated with these techniques.
数十年来的进化压力赋予了结核分枝杆菌（Mtb）极其复杂的生理和功能特性，使其成为数百年来感染人类的最可怕微生物之一。基因组学和基因组编辑工具取得的惊人进展极大地增进了我们对作为一种生物体的结核分枝杆菌的了解，但在将其作为病原体进行全面理解方面却遭遇了重大失败。尽管基于同源重组、反义技术、可控蛋白水解等的传统工具在推进我们对结核分枝杆菌病理生理学的理解方面做出了重要贡献，但由于某些技术限制，这些方法并未显著促进我们对分枝杆菌的探索。在这篇综述文章中，我们汇总了在分枝杆菌基因组编辑中实施的各种方法，以及成簇规律间隔短回文重复序列（CRISPR）干扰（CRISPRi）的最新应用，强调了与这些技术相关的成就和挑战。
