- 著者
- Huo Wenying Zhao Guannan Yin Jinggang Ouyang Xuan Wang Yinan Yang Chuanhe Wang Baojing Dong Peixin Wang Zhixiang Watari Hidemichi Chaum Edward Pfeffer Lawrence M. Yue Junming
- 出版者
- Ivyspring International Publisher
- 雑誌
- Journal of Cancer (ISSN:18379664)
- 巻号頁・発行日
- vol.8, no.1, pp.57-64, 2017
- 被引用文献数
- 81
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.
- 著者
- Yue Junming Du Ziyun Zhou Fu-Ming Dong Peixin Pfeffer Lawrence M
- 出版者
- OMICS International
- 雑誌
- Cancer Medicine & Anti Cancer Drugs
- 巻号頁・発行日
- vol.1, no.1, pp.1000103, 2016-03
The technology based on clustered regularly interspaced short palindromic repeats (CRISPR/Cas9) has been successfully applied to genome editing and has shown a promising future in gene functional studies. Human cancer is a complex disease due to multiple gene mutations, amplifications, deletions, up regulations or down regulations. It is a challenge to generate precise cell or animal cancer models in vitro and in vivo to investigate the complex process of cancer. The CRISPR/Cas9 technology provides a new opportunity to study human cancer by disrupting multiple genes or introducing point mutations at a specific locus of genome, and thus mimicking the features of human cancer in cell or animal models. Here we will review the current status of CRIPSR/Cas9 system and its potential application to cancer research.
- 著者
- Yue Junming Du Ziyun Zhou Fu-Ming Dong Peixin Pfeffer Lawrence M
- 出版者
- OMICS International
- 雑誌
- Cancer Medicine & Anti Cancer Drugs
- 巻号頁・発行日
- vol.1, no.1, 2016-03