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| Effect of Cas9 Protein on Biological and Ultrastructural Characteristics of the Human Bone-seeking Breast Cancer Cell Line |
| SONG Li-jie1,2, WANG Li2, YANG Chuan-hong2, LAI Huang-wen2, WANG Jie2 |
1 School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China;
2 Department of Medical Research, General Hospital of Guangzhou Military Command, Guangzhou 510010, China |
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Abstract Objective:To generate the human bone-seeking breast cancer cell line expressing Cas9 protein and study its biological and ultrastructural properties for the research of breast cancer cells' bone Metastasis using CRISPR-Cas9 technology. Methods:Cas9 protein gene was transfected into the human bone-seeking breast cancer cell line MDA-MB-231BO by lentiviral vector. After G418 screening, MDA-MB-231BO-Cas9 cell line which can constantly express Cas9 protein was obtained and examined by flow cytometry, Western blot and immunocytochemistry; Cell proliferation and migration were verified by real time cellular analysis and wound-healing assay; the effect of Cas9 expression on the ultrastructure of cells was observed under the transmission electron microscope. Results:The human bone-seeking breast cancer cell line MDA-MB-231BO-Cas9 expressing Cas9 protein has been constructed successfully, and the expression of Cas9 has no effect on cell's proliferation, migration and ultrastructure. Conclusion:MDA-MB-231BO-Cas9 cell line can be used for the further research using the CRISPR/Cas9 technology.
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Received: 21 January 2016
Published: 15 February 2016
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