Project/Area Number |
05454264
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Research Category |
Grant-in-Aid for General Scientific Research (B)
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Allocation Type | Single-year Grants |
Research Field |
Neurology
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Research Institution | Natinal Center of Neurology and Psychiatry |
Principal Investigator |
ARAHATA Kiichi (1994) Department of Neuromuscular Research, National Institute of Neuroscience, Director, 神経研究所・疾病研究第一部, 部長 (30053325)
杉田 秀夫 (1993) 国立精神・神経センター, 総長 (80009951)
|
Co-Investigator(Kenkyū-buntansha) |
TSUKAHARA Toshifumi Dept.of Neuromuscular Research, National Institute of Neuroscience, Investigator, 神経センター・神経研究所・疾病研究第一部, 研究員 (60207339)
TAKEDA Shinichi Dept.of Neuromuscular Research, National Institute of Neuroscience, Section Chie, 神経センター・神経研究所・疾病研究第一部, 室長 (90171644)
荒畑 喜一 神経研究所, 疾病研究第一部, 部長 (30053325)
|
Project Period (FY) |
1993 – 1994
|
Project Status |
Completed (Fiscal Year 1994)
|
Budget Amount *help |
¥7,700,000 (Direct Cost: ¥7,700,000)
Fiscal Year 1994: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1993: ¥5,100,000 (Direct Cost: ¥5,100,000)
|
Keywords | Gene transfer / Adenoviral vector / Cultured muscle cells / Regenerated muscle tissue / Muscular dystrophy / Gene therapy / 再生筋 / beta‐galactosidase(lacZ)遺伝子 / ジストロフィン遺伝子 / ミオシンH鎖IIBプロモーター / 正荷電リポソーム |
Research Abstract |
The effective gene transfer using adenoviral vector has been done in cultured muscle cells and in regenerated rat muscle tissue. 1) Gene transfer into cultured muscle cells We used two recombinant adeno viruses, Adex CA-1acZ,where 1ac Z was driven by ubiquitous strong promoter, and Adex MHC IIB-1acZ,where 1acZ was driven by muscle-specific promoter. When Adex CA 1acZ was transferred into mouse myogenic cell line, C2 cells under the condition of m.o.j 10,1ac Z was effectively transcribed and beta-galactosidase was apparent in all the cultured cells. When this recombinant adeno virus was infected into differentiated C2 cells, beta-galactosidase activities has been found even at large myotubes. This finding clearly showed that recombinant adeno virus can be transfected into post-mitotic cells. In the experiments with Adex MHC IIB-1acZ,beta-galactosidase activity was prominent only when recombinant adenovirus was infected into fully differentiated cells. This activation pattern reflects the character of MHC-IIB promoter, which was active in adult muscle tissue. 2) Gene transfer into the regenerated rat muscle tissue Adex CA-1ac Z has been injected into rat soleus muscle. Effective gene transfer has been detected, when muscle regeneration was induced by Bupivacaine treatment prior to recombinant adenoviral infection.
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