Gene targeting using the Agrobacterium tumefaciens-mediated CRISPR-Cas system in rice.
Gene targeting using the Agrobacterium tumefaciens-mediated CRISPR-Cas system in rice.
复制标题
在水稻中使用根癌农杆菌介导的 CRISPR-Cas 系统进行基因打靶。
DOI：
10.1186/s12284-014-0005-6
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Yang J
中科院分区：
文献类型：
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作者：
Xu R;Li H;Qin R;Wang L;Li L;Wei P;Yang J
II型簇状、规则间隔、短回文重复序列(CRISPR)/CRISPR相关蛋白9(Cas9)系统是一种新的位点特异性基因组修饰的分子工具。CRISPR-CAS9系统是最近通过瞬时或稳定转化引入植物的。在这里，我们报道了通过农杆菌介导的CRISPR-Cas9系统在水稻中进行基因打靶。设计了3个20个核苷酸的CRISPR RNA与不同的位点配对，然后是水稻除草剂抗性基因BEL的Protspacer邻近基序(PAM)。将单引导RNA(SgRNA)和Cas9盒整合到一个双元载体中，通过农杆菌介导法稳定转化获得了含有sgRNA：Cas9的转基因水稻植株。通过分析相应转基因植物基因组上的靶点，确定了突变。诱变率在~2%~~16%之间。表型分析表明，双等位突变的转基因植株对苯达松敏感。我们的结果表明，可以通过sgRNA：Cas9诱导的基因打靶来有目的地改变农业性状。CRISPR-CAS9系统可作为作物育种中性状改良的有力工具。
The type II clustered, regularly interspaced, short palindromic repeat (CRISPR)/ CRISPR-associated protein 9 (Cas9) system is a novel molecular tool for site-specific genome modification. The CRISPR-Cas9 system was recently introduced into plants by transient or stable transformation. Here, we report gene targeting in rice via the Agrobacterium tumefaciens-mediated CRISPR-Cas9 system. Three 20-nt CRISPR RNAs were designed to pair with diverse sites followed by the protospacer adjacent motif (PAM) of the rice herbicide resistance gene BEL. After integrating the single-guide RNA (sgRNA) and Cas9 cassette in a single binary vector, transgenic rice plants harboring sgRNA:Cas9 were generated by A. tumefaciens-mediated stable transformation. By analyzing the targeting site on the genome of corresponding transgenic plants, the mutations were determined. The mutagenesis efficiency was varied from ~2% to ~16%. Furthermore, phenotypic analysis revealed that the biallelic mutated transgenic plant was sensitive to bentazon. Our results indicate that the agricultural trait could be purposely modified by sgRNA:Cas9-induced gene targeting. CRISPR-Cas9 system could be exploited as a powerful tool for trait improvements in crop breeding.
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影响因子：
5.1
作者：
Belhaj K;Chaparro-Garcia A;Kamoun S;Nekrasov V
通讯作者：
Nekrasov V
DOI：
10.1126/science.1232033
发表时间：
2013-02-15
期刊：
Science (New York, N.Y.)
影响因子：
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作者：
Mali P;Yang L;Esvelt KM;Aach J;Guell M;DiCarlo JE;Norville JE;Church GM
通讯作者：
Church GM
影响因子：
7.4
作者：
Zhang, Yong;Zhang, Feng;Voytas, Daniel F.
通讯作者：
Voytas, Daniel F.
影响因子：
5.1
作者：
Pan, Gang;Zhang, Xianyin;Tu, Jumin
通讯作者：
Tu, Jumin
影响因子：
46.9
作者：
通讯作者：
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