| Project/Area Number |
17590425
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Virology
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| Research Institution | YOKOHAMA CITY UNIVERSITY SCHOOL OF MEDICINE |
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Principal Investigator |
SASAKI Shin YOKOHAMA CITY UNIV., SCH. OF MED, ASSOCIATE PROFESSOR, 医学部・客員準教授 (40342901)
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| Co-Investigator(Kenkyū-buntansha) |
TAKESHITA Fumihiko YOKOHAMA CITY UNIV., SCH. OF MED, ASSOCIATE PROFESSOR, 医学部, 準教授 (60333572)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2005: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | DNA VACCINE / TRANSCRUPTION FACTOR / GENE REGULATION |
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| Research Abstract |
MAINTAINANCE OF HIGH-LEVEL TRANSGENE EXPRESSION IS THE MAIN CHALLENGE IN CURRENT GENE THERAPY AND GENE-MEDIATED VACCINES. ALTHOUGH THE CYTOMEGALOVIRUS (CMV) PROMOTER/ENHANCER OR ITS DERIVATIVE THE CAG PROMOTER HAS BEEN HARNESSED IN CURRENT GENE THERAPY AND GENE-MEDIATED VACCINE VECTORS, TRANSGENE EXPRESSION BY THESE VECTORS IS OFTEN TRANSIENT AND REMAINS AT SUBOPTIMAL LEVELS DUE TO UNDEFINED MECHANISMS, POSSIBLY INCLUDING THE SHORTAGE OF TRANSCRIPTIONAL MACHINERY. TO OVERCOME THIS DRAWBACK, WE DESIGNED A NOVEL TRANSCRIPTIONAL CONTROL SYSTEM, DESIGNATED HERE AS TRANSCRIPTION FACTOR SUPERCHARGING PROMOTER SYSTEM, IN WHICH TRANSGENE EXPRESSION IS REGULATED BY THE POSITIVE FEEDBACK CIRCUIT CONSISTING OF CIS-AND TRANS-ACTING ELEMENTS OF GENE EXPRESSIONMACHINERY. AMONG COMBINATIOINS OF THESE ELEMENTS, A PLASMID COMPOSED OF A TARGET GENE EXPRESSION CASSETTE DRIVEN BY THE CHIMERIC CMV PROMOTER CONTAINING REPETITIVE 12-0-TETRADECANOYLPHORBOL-13-ACETATE-RESPONSIVE ELEMENTS AS CIS-ACTING ELEMENTS (CMV-TTT) AND EXPRESSION CASSETTES FOR C-FOS AND C-JUN GENES AS TRANS-ACTING ELEMENTS FACILITATED HIGH AND LONG-TERM (>10 MONTHS) EXPRESSION OF A TRANSGENE AFTER IYS INTRAMUSCULAR ELECTROPORATION-MEDIATED DELIVERY IN MICE.. SINCE HUMAN SECRETORY ALKALINE PHOSPHATASE WAS USED AS A REPORTER, IT WAS SUGGESTED THAT THE IMMUNE EVASION MECHANISM ELICITED BY THE CMV-TTT AND/OR C-FOS/C-JUN EXPRESSION ALSO CONTRIBUTED TO THE SUSTAINED EXPRESSION IN MICE.. OUR STRATEGY MAY OPEN A NEW AVENUE FOR A GENE THERAPY THAT INVOLVES LIFELONG SUPPLEMENTATION OF A DEFICIENT PROTEIN THAT COULD BE TARGETED THE HOST' S IMMUNE SYSTEM.
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