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Hokkaido University Collection of Scholarly and Academic Papers >
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Lentiviral CRISPR/Cas9 vector mediated miR-21 gene editing inhibits the epithelial to mesenchymal transition in ovarian cancer cells
This item is licensed under:Creative Commons Attribution-NonCommercial 4.0 International
| Title: | Lentiviral CRISPR/Cas9 vector mediated miR-21 gene editing inhibits the epithelial to mesenchymal transition in ovarian cancer cells |
| Authors: | Huo, Wenying Browse this author | | Zhao, Guannan Browse this author | | Yin, Jinggang Browse this author | | Ouyang, Xuan Browse this author | | Wang, Yinan Browse this author | | Yang, Chuanhe Browse this author | | Wang, Baojing Browse this author | | Dong, Peixin Browse this author →KAKEN DB | | Wang, Zhixiang Browse this author | | Watari, Hidemichi Browse this author →KAKEN DB | | Chaum, Edward Browse this author | | Pfeffer, Lawrence M. Browse this author | | Yue, Junming Browse this author |
| Keywords: | miR-21 | | CRISPR/Cas9 | | lentiviral vector | | ovarian cancer | | EMT |
| Issue Date: | 2017 |
| Publisher: | Ivyspring International Publisher |
| Journal Title: | Journal of Cancer |
| Volume: | 8 |
| Issue: | 1 |
| Start Page: | 57 |
| End Page: | 64 |
| Publisher DOI: | 10.7150/jca.16723 |
| Abstract: | CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats) mediated genome editing is a powerful approach for loss of function studies. Here we report that lentiviral CRISPR/Cas9 vectors are highly efficient in introducing mutations in the precursor miRNA sequence, thus leading to the loss of miRNA expression and function. We constructed four different lentiviral CRISPR/Cas9 vectors that target different regions of the precursor miR-21 sequence and found that these lentiviral CRISPR/Cas9 miR-21 gRNA vectors induced mutations in the precursor sequences as shown by DNA surveyor mutation assay and Sanger sequencing. Two miR-21 lentiviral CRISPR/Cas9 gRNA vectors were selected to probe miR-21 function in ovarian cancer SKOV3 and OVCAR3 cell lines. Our data demonstrate that disruption of pre-miR-21 sequences leads to reduced cell proliferation, migration and invasion. Moreover, CRISPR/Cas9-mediated miR-21 gene editing sensitizes both SKOV3 and OVCAR3 cells to chemotherapeutic drug treatment. Disruption of miR-21 leads to the inhibition of epithelial to mesenchymal transition (EMT) in both SKOV3 and OVCAR3 cells as evidenced by the upregulation of epithelial cell marker E-cadherin and downregulation of mesenchymal marker genes, vimentin and Snai2. The miR-21 target genes PDCD4 and SPRYT2 were upregulated in cells transduced with miR-21gRNAs compared to controls. Our study indicates that lentiviral CRISPR/Cas9-mediated miRNA gene editing is an effective approach to address miRNA function, and disruption of miR-21 inhibits EMT in ovarian cancer cells. |
| Rights: | https://creativecommons.org/licenses/by-nc/4.0/ |
| Type: | article |
| URI: | http://hdl.handle.net/2115/63900 |
| Appears in Collections: | 医学院・医学研究院 (Graduate School of Medicine / Faculty of Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 董 培新
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