| 通过CRISPR/Cas系统构建miR-17-92基因簇双切载体 |
| |
| 引用本文： | 金姝含,杜丽萍,邹肖肖,于泽,梁洋. 通过CRISPR/Cas系统构建miR-17-92基因簇双切载体[J]. 东北林业大学学报, 2017, 45(2) |
| |
| 作者姓名： | 金姝含 杜丽萍 邹肖肖 于泽 梁洋 |
| |
| 作者单位： | 东北林业大学,哈尔滨,150040 |
| |
| 基金项目： | 中央高校基本科研业务费项目，淡水鱼育种国家地方联合工程实验室开放课题，东北林业大学生命科学学院大学生创新训练项目 |
| |
| 摘 要： | 通过CRISPR/Cas系统来构建miR-17-92基因簇的双切载体,研究结果表明:在miR-17-92基因簇的序列上找到了2个PAM(TGG和GGG)位点,从而得到了2个原间隔序列,设计合成基因簇双链crRNA;合成该片段并将其插入到p X260载体,使得在同一个载体上虽然只有一个g-RNA,但能够同时切割两个不同的靶位点;将该载体转染NIH3T3细胞验证切割效率,PCR结果和测序结果显示,所构建的CRISPR系统可以对基因片段进行有效切割。
|
| 关 键 词： | CRISPR/Cas系统 miR-17-92基因簇 双切载体 crRNA |
Double-cut Carrier in Constructing miR-17-92 Gene Cluster by CRISPR/Cas9 System |
| |
| Jin Shuhan,Du Liping,Zhou Xiaoxiao,Yu Ze,LiangYang. Double-cut Carrier in Constructing miR-17-92 Gene Cluster by CRISPR/Cas9 System[J]. Journal of Northeast Forestry University, 2017, 45(2) |
| |
| Authors: | Jin Shuhan Du Liping Zhou Xiaoxiao Yu Ze LiangYang |
| |
| Abstract: | We constructed double-cut carrier of miR-17-92 gene cluster using CRISPR/Cas9 system.We identified two PAM and two protospacesare,and designed pre-crRNAs.The synthesized pre-crRNAs(that was the designed annealing oligonucleotides based on the sequence specific)were inserted between the pX260 carrier.Therefore,there is only one g-RNA in the same carrier,cutting two different target sites at the same time.NIH3T3 cells were transfected by plasmid of pX260 carrier including pre-crRNAs for the cutting effect.The miR-17-92 gene was cut by CRISPR/Cas system. |
| |
| Keywords: | CRISPR/Cas9 system miR-17-92 gene cluster Double-cut carrier crRNA |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
