Highly Efficient Targeted Gene Editing in Upland Cotton Using the CRISPR/Cas9 System.
Highly Efficient Targeted Gene Editing in Upland Cotton Using the CRISPR/Cas9 System.
复制标题
使用 CRISPR/Cas9 系统对陆地棉进行高效靶向基因编辑
DOI：
10.3390/ijms19103000
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发表时间：
2018-10-01
影响因子：
5.6
通讯作者：
Sun J
中科院分区：
文献类型：
--
作者：
Zhu S;Yu X;Li Y;Sun Y;Zhu Q;Sun J
成簇规则间隔短回文重复序列/CRISPR相关蛋白9（CRISPR/Cas9）基因编辑系统已被证明能够在包括棉花在内的许多植物的靶DNA中诱导高效诱变，并且已成为研究基因功能和作物改良的重要工具。在这里，我们开发了一种简单易操作的CRISPR/Cas9系统，并通过靶向ALARP（一种编码富含丙氨酸的蛋白质的基因，该蛋白质优先在棉纤维中表达）在棉花中展示了其高编辑效率。基于对10个转CRISPR/Cas9基因棉花靶位点的序列分析，我们发现GhALARP-A和GhALARP-D靶位点的突变频率分别为71.4-100%和92.9- 100%。最常见的编辑事件是删除，但也观察到删除和大插入。在大多数转基因植物中检测到镶嵌突变编辑事件。在分析的任何15个预测位点中均未检测到脱靶突变事件。本研究为进一步研究GhALARP在棉纤维发育中的作用提供了突变体。我们的研究结果进一步证明了CRISPR/Cas9作为棉花靶向诱变工具的可行性，为棉花靶向诱变和功能基因组学研究提供了有效的工具。
The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) gene editing system has been shown to be able to induce highly efficient mutagenesis in the targeted DNA of many plants, including cotton, and has become an important tool for investigation of gene function and crop improvement. Here, we developed a simple and easy to operate CRISPR/Cas9 system and demonstrated its high editing efficiency in cotton by targeting-ALARP, a gene encoding alanine-rich protein that is preferentially expressed in cotton fibers. Based on sequence analysis of the target site in the 10 transgenic cottons containing CRISPR/Cas9, we found that the mutation frequencies of GhALARP-A and GhALARP-D target sites were 71.4–100% and 92.9–100%, respectively. The most common editing event was deletion, but deletion together with large insertion was also observed. Mosaic mutation editing events were detected in most transgenic plants. No off-target mutation event was detected in any the 15 predicted sites analyzed. This study provided mutants for further study of the function of GhALARP in cotton fiber development. Our results further demonstrated the feasibility of use of CRISPR/Cas9 as a targeted mutagenesis tool in cotton, and provided an efficient tool for targeted mutagenesis and functional genomics in cotton.
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影响因子：
4.6
作者：
Chen X;Lu X;Shu N;Wang S;Wang J;Wang D;Guo L;Ye W
通讯作者：
Ye W
第一代杨树中 CRISPR/Cas9 介导的高效定点诱变
影响因子：
4.6
作者：
Fan D;Liu T;Li C;Jiao B;Li S;Hou Y;Luo K
通讯作者：
Luo K
影响因子：
17.3
作者：
Gaj, Thomas;Gersbach, Charles A.;Barbas, Carlos F., III
通讯作者：
Barbas, Carlos F., III
DOI：
10.1073/pnas.1019533108
发表时间：
2011-02-08
影响因子：
11.1
作者：
Mahfouz, Magdy M.;Li, Lixin;Zhu, Jian-Kang
通讯作者：
Zhu, Jian-Kang
影响因子：
27.5
作者：
Ma, Xingliang;Zhang, Qunyu;Liu, Yao-Guang
通讯作者：
Liu, Yao-Guang