Javed Muhammad R, Sadaf Maria, Ahmed Temoor, Jamil Amna, Nawaz Marium, Abbas Hira, Ijaz Anam
Department of Bioinformatics and Biotechnology, Government College University Faisalabad (GCUF), Faisalabad, 38000, Pakistan.
Curr Microbiol. 2018 Dec;75(12):1675-1683. doi: 10.1007/s00284-018-1547-4. Epub 2018 Aug 4.
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR or more precisely CRISPR-Cas) system has proven to be a highly efficient and simple tool for achieving site-specific genome modifications in comparison to Zinc Finger Nucleases (ZFNs) and Transcription Activator-Like Effector Nucleases (TALENs). The discovery of bacterial defense system that uses RNA-guided DNA cleaving enzymes for producing double-strand breaks along CRISPR has provided an exciting alternative to ZFNs and TALENs for gene editing & regulation, as the CRISPR-associated (Cas) proteins remain the same for different gene targets and only the short sequence of the guide RNA needs to be changed to redirect the site-specific cleavage. Therefore, in recent years the CRISPR-Cas system has emerged as a revolutionary engineering tool for carrying out precise and controlled genetic modifications in many microbes such as Escherichia coli, Staphylococcus aureus, Lactobacillus reuteri, Clostridium beijerinckii, Streptococcus pneumonia, and Saccharomyces cerevisiae. Though, concerns about CRISPR-Cas effectiveness in interlinked gene modifications and off-target effects need to be addressed. Nevertheless, it holds a great potential to speed up the pace of gene function discovery by interacting with previously intractable organisms and by raising the extent of genetic screens. Therefore, the potential applications of this system in microbial adaptive immune system, genome editing, gene regulations, functional genomics & biosynthesis along ethical issues, and possible harmful effects have been reviewed.
成簇规律间隔短回文重复序列（CRISPR，或更准确地说是CRISPR-Cas）系统已被证明是一种高效且简单的工具，与锌指核酸酶（ZFNs）和转录激活样效应核酸酶（TALENs）相比，可实现位点特异性基因组修饰。利用RNA引导的DNA切割酶沿CRISPR产生双链断裂的细菌防御系统的发现，为基因编辑和调控提供了一种令人兴奋的替代ZFNs和TALENs的方法，因为不同基因靶点的CRISPR相关（Cas）蛋白保持不变，只需改变引导RNA的短序列即可重定位位点特异性切割。因此，近年来，CRISPR-Cas系统已成为一种革命性的工程工具，可在许多微生物中进行精确且可控的基因修饰，如大肠杆菌、金黄色葡萄球菌、罗伊氏乳杆菌、拜氏梭菌、肺炎链球菌和酿酒酵母。不过，CRISPR-Cas在连锁基因修饰中的有效性和脱靶效应等问题仍需解决。尽管如此，它在与以前难以处理的生物体相互作用以及扩大遗传筛选范围方面，具有加快基因功能发现步伐的巨大潜力。因此，本文综述了该系统在微生物适应性免疫系统、基因组编辑、基因调控、功能基因组学和生物合成中的潜在应用，以及相关伦理问题和可能的有害影响。
