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CRISPR-Cas介导的基因组编辑:遗传操作领域的重大变革

Genome Editing by CRISPR-Cas: A Game Change in the Genetic Manipulation of .

作者信息

Ghribi Manel, Nouemssi Serge Basile, Meddeb-Mouelhi Fatma, Desgagné-Penix Isabel

机构信息

Department of Chemistry, Biochemistry and Physics, Université du Québec à Trois-Rivières, 3351, Boulevard des Forges, C.P. 500, Trois-Rivières, QC G9A 5H7, Canada.

Groupe de Recherche en Biologie Végétale, Université du Québec à Trois-Rivières, 3351, Boulevard des Forges, C.P. 500, Trois-Rivières, QC G9A 5H7, Canada.

出版信息

Life (Basel). 2020 Nov 20;10(11):295. doi: 10.3390/life10110295.

Abstract

Microalgae are promising photosynthetic unicellular eukaryotes among the most abundant on the planet and are considered as alternative sustainable resources for various industrial applications. is an emerging model for microalgae to be manipulated by multiple biotechnological tools in order to produce high-value bioproducts such as biofuels, bioactive peptides, pigments, nutraceuticals, and medicines. Specifically, has become a subject of different genetic-editing techniques adapted to modulate the production of microalgal metabolites. The main nuclear genome-editing tools available today include zinc finger nucleases (ZFNs), transcriptional activator-like effector nucleases (TALENs), and more recently discovered the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein (Cas) nuclease system. The latter, shown to have an interesting editing capacity, has become an essential tool for genome editing. In this review, we highlight the available literature on the methods and the applications of CRISPR-Cas for genetic engineering, including recent transformation methods, most used bioinformatic tools, best strategies for the expression of Cas protein and sgRNA, the CRISPR-Cas mediated gene knock-in/knock-out strategies, and finally the literature related to CRISPR expression and modification approaches.

摘要

微藻是地球上最丰富的光合单细胞真核生物之一,被视为各种工业应用中可持续的替代资源。为了生产生物燃料、生物活性肽、色素、营养保健品和药物等高价值生物产品,微藻已成为多种生物技术工具可操控的新兴模式生物。具体而言,微藻已成为不同基因编辑技术的研究对象,这些技术用于调节微藻代谢产物的产生。目前可用的主要核基因组编辑工具包括锌指核酸酶(ZFNs)、转录激活样效应因子核酸酶(TALENs),以及最近发现的成簇规律间隔短回文重复序列(CRISPR)-CRISPR相关蛋白(Cas)核酸酶系统。后者具有有趣的编辑能力,已成为基因组编辑的重要工具。在本综述中,我们重点介绍了关于CRISPR-Cas在微藻基因工程中的方法和应用的现有文献,包括最近的转化方法、最常用的生物信息学工具、Cas蛋白和sgRNA表达的最佳策略、CRISPR-Cas介导的基因敲入/敲除策略,以及最后与CRISPR表达和修饰方法相关的文献。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f806/7699682/cde40f0fdc7f/life-10-00295-g001.jpg

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