State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, 214122, China.
School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, 214122, China.
Appl Microbiol Biotechnol. 2017 Nov;101(22):8063-8075. doi: 10.1007/s00253-017-8486-z. Epub 2017 Sep 30.
Functional genomics of filamentous fungi has gradually uncovered gene information for constructing 'cell factories' and controlling pathogens. Available gene manipulation methods of filamentous fungi include random integration methods, gene targeting technology, gene editing with artificial nucleases and RNA technology. This review describes random gene integration constructed by restriction enzyme-mediated integration (REMI); Agrobacterium-mediated transformation (AMT); transposon-arrayed gene knockout (TAGKO); gene targeting technology, mainly about homologous recombination; and modern gene editing strategies containing transcription activator-like effector nucleases (TALENs) and a clustered regularly interspaced short palindromic repeat/associated protein system (CRISPR/Cas) developed in filamentous fungi and RNA technology including RNA interference (RNAi) and ribozymes. This review describes historical and modern gene manipulation methods in filamentous fungi and presents the molecular tools available to researchers investigating filamentous fungi. The biggest difference of this review from the previous ones is the addition of successful application and details of the promising gene editing tool CRISPR/Cas9 system in filamentous fungi.
丝状真菌的功能基因组学逐渐揭示了构建“细胞工厂”和控制病原体的基因信息。丝状真菌的现有基因操作方法包括随机整合方法、基因靶向技术、人工核酸酶介导的基因编辑和 RNA 技术。本文综述了限制性内切酶介导的整合(REMI)构建的随机基因整合;农杆菌介导的转化(AMT);转座子排列基因敲除(TAGKO);基因靶向技术,主要是同源重组;以及丝状真菌中开发的现代基因编辑策略,包括转录激活因子样效应物核酸酶(TALENs)和成簇规律间隔短回文重复/相关蛋白系统(CRISPR/Cas)和 RNA 技术,包括 RNA 干扰(RNAi)和核酶。本文综述了丝状真菌中历史和现代的基因操作方法,并介绍了研究丝状真菌的研究人员可用的分子工具。与之前的综述相比,本文最大的不同之处在于增加了 CRISPR/Cas9 系统这一有前途的基因编辑工具在丝状真菌中的成功应用和详细信息。