| Project/Area Number |
09309011
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
広領域
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| Research Institution | Chiba Institute of Technology |
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Principal Investigator |
TAKAKU Hiroshi Chiba Institute of Technology, Facutly of Technology, Professor, 工学部, 教授 (50101267)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAI Kazuyuki Chiba Institute of Technology, Facuqy of Technology, Lecturer, 工学部, 講師 (40260848)
YAMAMOTO Naoki Tokyo Medical and Dental University, Medical Facutly, Professor, 医学部, 教授 (00094053)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥28,200,000 (Direct Cost: ¥28,200,000)
Fiscal Year 1999: ¥5,400,000 (Direct Cost: ¥5,400,000)
Fiscal Year 1998: ¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 1997: ¥17,600,000 (Direct Cost: ¥17,600,000)
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| Keywords | circular dumbbell RNA / DNA chimeric oligonucleotide / antisense DNA / sense RNA / Rnase H / retrovirus / HIV-1 infected cells / nuclease resistance / RNase H活性 / 耐分解酵素性 |
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| Research Abstract |
We have designed a new class of oligonucleotides, "dumbbell RNA/DNA chimeric phosphodiesters", containing two alkyl loop structures with RNA/DNA base pairs (sense (RNA) and antisense (DNA)) in the double helical stem. The reaction of nicked (NDRDON-gag-AUG) and circular (CDRDON-gag-AUG) dumbbell RNA/DNA chimeric oligonucleotides with RNaseH gave the corresponding antisense phosphodiester oligonucleotide together with the sense RNA cleavage products. The liberated antisense phosphodiester oligodeoxynucleotide was bound to the target RNA. The circular dumbbell RNA/DNA chimeric oligoncleotide showed more nuclease resistance and cellular uptake than the linear antisense phosphorothioate oligodeoxynucleotide (S-ODN-gag-AUG) and nicked dumbbell RNA/DNA chimeric oligonucleotide. The CDRDON-gag -AUG with an AUG initiation codon sequence, as the target of the HIV-1 gag-gene (779-801), was synthesized and tested for inhibitory effects using PBMCs. The circular dumbbell RNA/DNA chimeric oligonucleotide(CDRDON-gag-AUG) showed highly inhibitory effects as compared to the antisense phosphorothioate oligonucleotide (S-ODN-gag-AUG), indicating sequence-specific inhibition of HIV-1 replication without the inhibition of reverse transcriptase and/or the viral entry process such as antisense phosphorothioate oligonucleotides.
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