2018 Fiscal Year Annual Research Report
ＣＲＩＳＰＲ／Ｃａｓ９封入高分子集合体の構築と生体内デリバリーへの挑戦
| Project/Area Number |
16F16355
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| Research Institution | The University of Tokyo |
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Principal Investigator |
宮田 完二郎 東京大学, 大学院工学系研究科(工学部), 准教授 (50436523)
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| Co-Investigator(Kenkyū-buntansha) |
MIN HYUN SU 東京大学, 工学(系)研究科(研究院), 外国人特別研究員
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| Project Period (FY) |
2016-10-07 – 2019-03-31
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| Keywords | Cas9 / messenger RNA delivery / polyplex |
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| Outline of Annual Research Achievements |
Cas9 activities of various cationic polymers were evaluated by luciferase-based Cas9 activity system. This assay consists of three different plasmids - sgRNA-expressing plasmid, Stop codon-contained luciferase-expressing plasmid, and luciferase donor DNA plasmid, which were kindly donated by Professor Moritoshi Sato (UTokyo). A cationic polymer (PAsp(OCT/DET)) was complexed with Cas9 messenger RNA and then the resulting polyplexes were transfected into human cervical cancer (HeLa) cells. The Cas9 activities were measured by a luminescence microplate reader. The cationic polymer elicited the similar luminescence intensity compared to commercially available transfection agent, lipofectamine 3000. This result indicates that the new cationic polymer is promising for in vitro Cas9 mRNA transfection.
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| Research Progress Status |
平成30年度が最終年度であるため、記入しない。
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| Strategy for Future Research Activity |
平成30年度が最終年度であるため、記入しない。
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