Department of Zoology, Biomedical Technology & Human Genetics, University School of Sciences, Gujarat University, Ahmedabad, Gujarat, India; PanGenomics International Pvt Ltd, Sterling Accuris Diagnostics, Ellis Bridge, Ahmedabad, Gujarat, India.
Rivaara Labs Pvt Ltd, KD Hospital, Vaishnodevi Circle, Ahmedabad, Gujarat, India.
Prog Mol Biol Transl Sci. 2021;180:69-84. doi: 10.1016/bs.pmbts.2021.01.005. Epub 2021 Mar 12.
CRISPR/Cas9, once discovered as an adaptive immune system in bacteria, has emerged as a disruptive technology in the field of genetic engineering. Technological advancements in the recent past has enhanced the applicability of CRISPR/Cas9 tool for gene editing, gene therapies, developmental studies and mutational analysis in various model organisms. Zebrafish, one of the excellent animal models, is preferred for conducting CRISPR/Cas9 studies to assess the functional implication of specific genes of interest. CRISPR/Cas9 mediated gene editing techniques, such as, knock-out and knock-in approaches, provide evidences to identify the role of different genes through loss-of-function studies. Also, CRISPR/Cas9 has been proved to be an efficient tool for designing disease models for gene expression studies based on phenotypic screening. The present chapter provides an overview of CRISPR/Cas9 mechanism, different strategies for DNA modifications and gene function analysis, highlighting the translational applications for future prospects, such as screening of drug toxicity and efficacy.
CRISPR/Cas9 最初在细菌中被发现是一种适应性免疫系统，现已成为基因工程领域的颠覆性技术。最近的技术进步增强了 CRISPR/Cas9 工具在基因编辑、基因治疗、发育研究和各种模式生物中的突变分析中的适用性。斑马鱼是优秀的动物模型之一，常用于进行 CRISPR/Cas9 研究，以评估特定感兴趣基因的功能意义。CRISPR/Cas9 介导的基因编辑技术，如敲除和敲入方法，通过功能丧失研究提供了识别不同基因作用的证据。此外，CRISPR/Cas9 已被证明是一种有效的工具，可用于基于表型筛选设计用于基因表达研究的疾病模型。本章概述了 CRISPR/Cas9 的机制、用于 DNA 修饰和基因功能分析的不同策略，强调了其未来的转化应用前景，如药物毒性和疗效的筛选。
