2005 Fiscal Year Final Research Report Summary
Functionalization of oligonucleotides capable to form alternate-stranded triplex containing stereoisomeric nucleotide units
| Project/Area Number |
15550144
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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 |
Chemistry related to living body
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| Research Institution | Gunma University |
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Principal Investigator |
SHINOZUKA Kazuo GUNMA UNIVERSITY, Chemistry, Professor, 工学部, 教授 (20206105)
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| Co-Investigator(Kenkyū-buntansha) |
MORIGUCHI Tomohisa GUNMA UNIVERSITY, Chemistry, Associate Professor, 工学部, 助教授 (70334133)
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| Project Period (FY) |
2003 – 2005
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| Keywords | α-β chimeric oligoDNA / intercalator / alternate-stranded triplex / novel non-nucleosidic component / diastereomer / pyrimidine-gap / metal complex / functional chimera DNA |
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| Research Abstract |
A novel phosphoramidite derivative of a dinucleoside phosphotriester unit bearing an intercalative moiety at its internucleotide linkage in a stereospecific manner was prepared and successfully incorporated into the middle portion of α-β chimeric oligoDNA. One of the resulting stereoisomeric chimera DNAs strongly enhances the thermal stability of an alternate-stranded triplex formed between the chimera and a double-stranded DNA. On the other hand, the duplexes containing the oligomers bearing the same unit failed to show any Tm increment (stabilization) effect regardless of the stereochemistry of the phosphorous atom bearing the intercalative moiety. Thus, the stabilizing effect of the current dinucleoside unit seems to be specific to the DNA triplex.
An anthraquinone derivative conjugated with 2,2-bis(hydroxymethyl)propionic acid as a novel non-nucleosidic component was synthesized and successfully incorporated into an internal region of 14-mer triplex-forming oligonucleotide (TFO) via
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the phosphoramidite method. The resulted novel TFO exhibits remarkable enhancement effect on the thermal stability of a DNA triplex upon binding to a pyrimidine-gap containing polypurine sequence.
Novel α-β chimeric oligodeoxynucleotides bearing an intercalator-conjugated nucleobase located at the internal 4-nt linker region were synthesized, and their triplex-stabilizing property was examined. The triple helical DNA formed between the modified chimera DNA and double-stranded DNA exhibited remarkable thermal stability ; however, the position of the intercalator-conjugated nucleobase had little influence on the stability. Among the examined, modified chimera DNA bearing the two intercalator-conjugated nucleobases at adjacent positions exhibited the highest stability.
Novel diastereomeric α-β chimeric oligodeoxynucleotides bearing a propionic acid derivative of an anthraquinone-polyamine conjugate in the "linker" region were synthesized. Upon hybridization with double-stranded DNA, the modified chimera DNAs formed substantially stable alternate-stranded triplex even at near physiological pH. Furthermore, the triplex-stabilizing effect brought about by the incorporation of the derivative was almost regardless of the stereochemistry of the derivative.
Novel conjugate composed of an intercalator-metal ligand-2,2-bis(hydroxymethyl)propionic acid was prepared. The conjugate forms complexes with divalent metal ions such as Fe^<2+>, Cu^<2+>, Zn^<2+>, etc. Among them, the Fe^<2+> complex cleaves double-stranded DNA, efficiently. A study to utilize a functional chimera DNA bearing the complex as a sequence specific gene regulator is now under way. Less
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