2016 Fiscal Year Final Research Report
Elucidation of SOX2 transcriptional regulation mechanism using genome editing technique
| Project/Area Number |
26460465
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Experimental pathology
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| Research Institution | Shimane University |
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Principal Investigator |
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| Co-Investigator(Renkei-kenkyūsha) |
KURAMITU Yasuhiro 山口大学, 医学部, 准教授 (50281811)
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| Project Period (FY) |
2014-04-01 – 2017-03-31
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| Keywords | ゲノム編集 / 幹細胞 |
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| Outline of Final Research Achievements |
Cas9, gRNA expression vectors, and homologous recombination vector were transfected into breast cancer cell line MCF-7 using genome editing. fluorescent genes Venus was inserted into the C-terminal side of the SOX2 gene. In the cell line of the homologous recombinant, although it localized in the nucleus and emitted green fluorescence, the fluorescence was weak, and it was difficult to analyze the localization of intracellular cells as live cells by fluorescence microscopy. In addition, there was variation in cell proliferation for each cell line compared to the parent strain, and off - target by gRNA was predicted. Similar results were found in genomic editing of other genes. Therefore, Cas9 protein and gRNA complex were formed in vitro and introduced. Currently, we analyze new homologous recombinants.
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| Free Research Field |
腫瘍学 RNA学
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