Genome editing in Ustilago maydis using the CRISPR-Cas system
Genome editing in Ustilago maydis using the CRISPR-Cas system
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DOI：
10.1016/j.fgb.2015.09.001
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发表时间：
2016-04-01
影响因子：
3
通讯作者：
Kahmann, Regine
中科院分区：
文献类型：
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作者：
Schuster, Mariana;Schweizer, Gabriel;Kahmann, Regine
本通讯描述了建立II型细菌CRISPR-CAS9系统，以有效干扰玉米真菌病原菌Ustilomaydis中的靶基因。在U6单链RNA启动子的控制下，一步转化表达UMAYDIS密码子优化的Cas9基因和合适的sgRNA的自复制质粒就足以诱导基因组编辑。平均而言，单个转化子的后代中有70%在各自的b基因内被破坏。在没有选择的情况下，自我复制的质粒会迅速丢失，从而允许CRISPR-Cas9系统的瞬时表达，从而将Cas9的潜在长期负面影响降至最低。这项技术将是功能冗余基因和基因家族同时中断的重要进展，以研究它们对玉米黑粉菌毒力的贡献。(C)2015 Elsevier Inc.保留所有权利。
This communication describes the establishment of the type II bacterial CRISPR-Cas9 system to efficiently disrupt target genes in the fungal maize pathogen Ustilogo maydis. A single step transformation of a self-replicating plasmid constitutively expressing the U. maydis codon-optimized cas9 gene and a suitable sgRNA under control of the U. maydis U6 snRNA promoter was sufficient to induce genome editing. On average 70% of the progeny of a single transformant were disrupted within the respective b gene. Without selection the self-replicating plasmid was lost rapidly allowing transient expression of the CRISPR-Cas9 system to minimize potential long-term negative effects of Cas9. This technology will be an important advance for the simultaneous disruption of functionally redundant genes and gene families to investigate their contribution to virulence of U. maydis. (C) 2015 Elsevier Inc. All rights reserved.