CRISPR/Cas9-Based Editing of Streptomyces for Discovery, Characterization, and Production of Natural Products.
CRISPR/Cas9-Based Editing of Streptomyces for Discovery, Characterization, and Production of Natural Products.
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基于 CRISPR/Cas9 的链霉菌编辑用于天然产物的发现、表征和生产
DOI：
10.3389/fmicb.2018.01660
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发表时间：
2018
影响因子：
5.2
通讯作者：
Sun Y
中科院分区：
文献类型：
--
作者：
Tao W;Yang A;Deng Z;Sun Y
微生物天然产物（NPs），特别是链霉菌属，已被认为是药物发现的一个无与伦比的资源。此外，测序技术和计算资源的最新进展进一步加强了从链霉菌基因组中鉴定大量NP生物合成基因簇（BGCs）的能力。然而，这些BGCs中的大多数在本地菌株中沉默或表达不良，仍然需要激活和研究，这在很大程度上依赖于有效的基因组编辑方法。因此，开发了许多策略，特别是最近开发的群集规则间隔短回文重复序列(CRISPR)/CRISPR相关（Cas）系统显示出包括链霉菌在内的各种模式生物的基因组编辑具有显着的更高准确性和效率。在这篇综述中，我们重点介绍了基于CRISPR/ cas9的方法在链霉菌中的应用，重点介绍了体内或体外BGCs的编辑，以及大尺寸BGCs的靶向克隆和在遗传可操纵宿主中的异源表达，用于发现、表征、重组和潜在药物的生产。
Microbial natural products (NPs) especially of the Streptomyces genus have been regarded as an unparalleled resource for pharmaceutical drugs discovery. Moreover, recent progress in sequencing technologies and computational resources further reinforces to identify numerous NP biosynthetic gene clusters (BGCs) from the genomes of Streptomyces. However, the majority of these BGCs are silent or poorly expressed in native strains and remain to be activated and investigated, which relies heavily on efficient genome editing approaches. Accordingly, numerous strategies are developed, especially, the most recently developed, namely, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated (Cas) system reveals remarkable higher accuracy and efficiency for genome editing in various model organisms including the Streptomyces. In this mini review, we highlight the application of CRISPR/Cas9-based approaches in Streptomyces, focus on the editing of BGCs either in vivo or in vitro, as well as target cloning of large-sized BGCs and heterologous expression in a genetically manipulatable host, for discovery, characterization, reengineering, and production of potential pharmaceutical drugs.