Japan Science and Technology Agency

Tokyo Institute of Technology

Akihiro Ohkubo

Kohji Seio

Mitsuo Sekine

Takeshi Sasami

Kazushi Sakamoto

Methods of synthesizing nucleic acid oligomers on a solid-phase support having a 3'-end nucleoside unit introduced thereon-as represented by formula II: wherein of formula II represents a 2&#x2032;-deoxyribonucleoside or its N-protected derivative, the substituent &#x2014;O&#x2014;(R)Si(R)&#x2014;(CHR)&#x2014;(CH)&#x2014;O&#x2014;P(OR)XO)&#x2014;(CH)is attached at the 3&#x2032; position of the sugar moiety of the nucleoside substituent; each of Rand Ris an alkyl or optionally substituted aryl group, wherein the optionally substituted aryl group has a substituent selected from the group consisting of Calkyl, nitro, cyano, halo and methoxyl; Ris a protecting group; X is S or O; Ris H or 4,4&#x2032;-dimethoxytrityl; each n is an integer of from 1 to 5; and the solid-phase support has hydroxyl groups on its surface.

3'-end nucleoside unit comprising phosphoramidite

To provide an oligonucleotide derivative that can be used without a problem of falling of a DNA probe from a support. The oligonucleotide derivative of the present invention is represented by the following General Formula (1). The oligonucleotide derivative of the present invention can be used in a DNA chip, a microarray, and so on and further used in a method of detecting gene and a method of regulating gene expression, for example. 

Oligonucleotide derivative, probe for detection of gene, and DNA chip

A phosphoramidite method for the synthesis of a nucleic acid oligomer without protecting the base moiety characterized in that a 3' or 5&#x2032; hydroxyl group of a nucleotide is reacted with a nucleoside phosphoramidite, a cyclonucleoside phosphoramidite, a 2&#x2032;-substituted nucleoside phosphoramidite, a 4&#x2032;-substituted nucleoside phosphoramidite, or a 2&#x2032;,4&#x2032;-di-substituted nucleoside phosphoramidite to produce a phosphodiester linkage. The phosphoramidite is contacted with an activator containing both a) hydroxybenzotriazole-1-ol (HOBt), a mono-substituted HOBt, a di-substituted HOBt, or a di-substituted phenol and b) imidazole, tetrazole, benzimidazoletriflate (BIT), 4-ethylthiotetrazole, imidazolium triflate(trifluoromethane sulfonate) or 4,5-dicyanoimidazole.

Method for the synthesis of nucleic acid without protecting base moiety

The purpose of the invention is to develop a silyl linker that can be efficiently introduced on a solid-phase support used for the synthesis of nucleic acid oligomers such as DNA. The present invention relates to a silyl linker for use in the solid-phase synthesis of nucleic acid, comprised of a compound of the general formula or its ester or salt:H&#x2014;(R1)Si(R2)-(CH)&#x2014;CONH-(A)-COOH&#x2003;&#x2003;(I)wherein each of R1 and R2 is an alkyl or aryl group, and (A) represents a spacer moiety; a 3'-end nucleoside unit having said compound linked via an oxygen atom to the 3-position of a sugar of the nucleoside or its derivative, a solid-phase support having the 3&#x2032;-end nucleoside unit, and a method for synthesis of nucleic acid oligomer with the use of said solid-phase support.

Growing community of inventors

Machida, Japan

Akihiro Ohkubo