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丝状真菌和卵菌中的 CRISPR-Cas9 基因组编辑方法。

CRISPR-Cas9 genome editing approaches in filamentous fungi and oomycetes.

机构信息

Max Planck Institute for Terrestrial Microbiology, Dept. Organismic Interactions, 35043 Marburg, Germany.

出版信息

Fungal Genet Biol. 2019 Sep;130:43-53. doi: 10.1016/j.fgb.2019.04.016. Epub 2019 Apr 29.

Abstract

Due to their biotechnological relevance as well as their importance as disease agents, filamentous fungi and oomycetes have been prime candidates for genetic selection and in vitro manipulation for decades. With the advent of new genome editing technologies such manipulations have reached a new level of speed and sophistication. The CRISPR-Cas9 genome editing technology in particular has revolutionized the ways how desired mutations can be introduced. To date, the CRISPR-Cas9 genome editing system has been established in more than 40 different species of filamentous fungi and oomycetes. In this review we describe the various approaches taken to assure expression of the components necessary for editing and describe the varying strategies used to achieve gene disruptions, gene replacements and precise editing. We discuss potential problems faced when establishing the system, propose ways to circumvent them and suggest future approaches not yet realized in filamentous fungi or oomycetes.

摘要

由于丝状真菌和卵菌在生物技术方面的重要性以及它们作为疾病因子的重要性,几十年来,它们一直是遗传选择和体外操作的主要候选对象。随着新的基因组编辑技术的出现,这些操作已经达到了新的速度和复杂性水平。特别是 CRISPR-Cas9 基因组编辑技术彻底改变了引入所需突变的方式。迄今为止,CRISPR-Cas9 基因组编辑系统已经在 40 多种不同的丝状真菌和卵菌中建立。在这篇综述中,我们描述了为确保编辑所需成分的表达而采取的各种方法,并描述了实现基因缺失、基因替换和精确编辑的各种策略。我们讨论了在建立该系统时面临的潜在问题,提出了克服这些问题的方法,并提出了尚未在丝状真菌或卵菌中实现的未来方法。

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