Nargesi Sanaz, Kaboli Saeed, Thekkiniath Jose, Heidari Somayeh, Keramati Fatemeh, Seyedmousavi Seyedmojtaba, Hedayati Mohammad Taghi
Department of Medical Mycology, School of Medicine, Mazandaran University of Medical Sciences, Sari 481751665, Iran.
Department of Medical Biotechnology, School of Medicine, Zanjan University of Medical Sciences, Zanjan 4513956111, Iran.
J Fungi (Basel). 2021 Mar 30;7(4):257. doi: 10.3390/jof7040257.
Manipulating fungal genomes is an important tool to understand the function of target genes, pathobiology of fungal infections, virulence potential, and pathogenicity of medically important fungi, and to develop novel diagnostics and therapeutic targets. Here, we provide an overview of recent advances in genetic manipulation techniques used in the field of medical mycology. Fungi use several strategies to cope with stress and adapt themselves against environmental effectors. For instance, mutations in the 14 alpha-demethylase gene may result in azole resistance in strains and shield them against fungicide's effects. Over the past few decades, several genome editing methods have been introduced for genetic manipulations in pathogenic fungi. Application of restriction enzymes to target and cut a double-stranded DNA in a pre-defined sequence was the first technique used for cloning in and . Genome editing technologies, including zinc-finger nucleases (ZFNs) and transcriptional activator-like effector nucleases (TALENs), have been also used to engineer a double-stranded DNA molecule. As a result, TALENs were considered more practical to identify single nucleotide polymorphisms. Recently, Class 2 type II Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas9 technology has emerged as a more useful tool for genome manipulation in fungal research.
操纵真菌基因组是了解目标基因功能、真菌感染的病理生物学、致病潜力以及医学上重要真菌的致病性,以及开发新型诊断和治疗靶点的重要工具。在此,我们概述了医学真菌学领域中使用的基因操纵技术的最新进展。真菌采用多种策略来应对压力并使自身适应环境效应物。例如,14α-脱甲基酶基因突变可能导致菌株产生唑抗性并使其免受杀菌剂的影响。在过去几十年中,已经引入了几种基因组编辑方法用于致病真菌的基因操纵。应用限制性内切酶以预定义序列靶向并切割双链DNA是最早用于克隆的技术。基因组编辑技术,包括锌指核酸酶(ZFNs)和转录激活样效应物核酸酶(TALENs),也已用于构建双链DNA分子。因此,TALENs被认为在识别单核苷酸多态性方面更实用。最近,II类2型成簇规律间隔短回文重复序列(CRISPR)/Cas9技术已成为真菌研究中基因组操纵的更有用工具。