Wege Sarah-Maria, Gejer Katharina, Becker Fabienne, Bölker Michael, Freitag Johannes, Sandrock Björn
Department of Biology, Philipps-University Marburg, 35037 Marburg, Germany.
Department of Physics, Philipps-University Marburg, 35037 Marburg, Germany.
J Fungi (Basel). 2021 Feb 18;7(2):149. doi: 10.3390/jof7020149.
The phytopathogenic smut fungus is a versatile model organism to study plant pathology, fungal genetics, and molecular cell biology. Here, we report several strategies to manipulate the genome of by the CRISPR/Cas9 technology. These include targeted gene deletion via homologous recombination of short double-stranded oligonucleotides, introduction of point mutations, heterologous complementation at the genomic locus, and endogenous N-terminal tagging with the fluorescent protein mCherry. All applications are independent of a permanent selectable marker and only require transient expression of the endonuclease Cas9hf and sgRNA. The techniques presented here are likely to accelerate research in the community but can also act as a template for genome editing in other important fungi.
植物病原性黑粉菌是研究植物病理学、真菌遗传学和分子细胞生物学的一种通用模式生物。在此，我们报告了几种利用CRISPR/Cas9技术操纵其基因组的策略。这些策略包括通过短双链寡核苷酸的同源重组进行靶向基因缺失、引入点突变、在基因组位点进行异源互补以及用荧光蛋白mCherry进行内源性N端标记。所有应用均不依赖于永久选择标记，仅需要内切核酸酶Cas9hf和sgRNA的瞬时表达。本文介绍的技术可能会加速该领域的研究，但也可作为其他重要真菌基因组编辑的模板。
