Functionalization of oligonucleotides capable to form alternate-stranded triplex containing stereoisomeric nucleotide units

能够形成含有立体异构核苷酸单元的交替链三链体的寡核苷酸的功能化

基本信息

批准号：
15550144
负责人：
SHINOZUKA Kazuo
金额：
$ 2.37万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

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 … More 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

制备了一个新型的二核苷酸磷酸三位酯单位的磷光衍生物，该单元在其插室部分以立体特异性的方式构成了嵌入式部分，并成功地掺入了α-β嵌合寡头的中部。所得的立体异构体Chimera DNA强烈增强了嵌合体和双链DNA之间形成的交替链三重的热稳定性。另一方面，双链体包含具有相同单位的低聚物，无论带有插入性部分的磷原子的立体化学，无论是磷原子的立体化学而言，都无法显示任何TM增量（稳定）效应。这是，当前的二核苷单元的稳定作用似乎特定于DNA三倍体。与2,2-双（羟甲基）丙酸连接的蒽醌衍生物作为一种新型的非核糖苷成分，并成功地纳入了14米型oligormenamemig offortirortirortient（均为14兆）的区域（方法。 The resulted novel TFO exhibits remarkable enhancement effect on the thermal stability of a DNA triple upon binding to a pyrimidine-gap containing polypurine sequence.Novel α-β chimeric oligodeoxynucleotides bearing an intercalator-conjugated nuclearbase located at the internal 4-nt linker region were synthesized, and their triple-stabilizing property was examined.在修饰的嵌合体DNA和双链DNA之间形成的三重螺旋DNA暴露于显着的热稳定性。然而，嵌入量偶联的核酶的位置对稳定性影响不大。在相邻位置的两个嵌入量偶联的核酶的经过的经过修饰的嵌合DNA中，在暴露于最高稳定性的位置。成initoseomericα-β嵌合寡脱氧核苷酸含有丙酰胺 - 喹酮 - 丙烯胺偶联物在链酸酯烯丙基二核苷酸衍生物中的丙二酸衍生物，在“链酸酯烯酮固定胺”中是“链接区域”的区域。在与双链DNA杂交后，修饰的嵌合体DNA甚至在近乎物理pH下也形成了基本稳定的替代链三元。此外，无论衍生物的立体化学何种衍生物的融合而产生的三重稳定效应。共轭形成二价金属离子，例如fe^<2+>，cu^<2+>，zn^<2+>等。其中，有效地，fe^<2+>复合物裂解了双链DNA。现在正在进行一项利用具有序列特定基因调节因子的功能性嵌合体DNA。较少的