Inhibition of HIV-1 replication of an HIV-1 dependent antisense oligonucleotides.

HIV-1 依赖性反义寡核苷酸抑制 HIV-1 复制。

• 批准号: 09309011
• 负责人: TAKAKU Hiroshi
• 金额: $ 18.05万
• 依托单位: Chiba Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09309011/
• 关键词: 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

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.

我们设计了一类新的寡核苷酸，“哑铃状RNA/DNA嵌合磷酸二酯”，在双螺旋茎中含有两个具有RNA/DNA碱基对（正义（RNA）和反义（DNA））的烷基环结构。缺口（NDRDON-gag-AUG）和环状（CDRDON-gag-AUG）哑铃状RNA/DNA嵌合寡核苷酸与RNaseH反应，产生相应的反义磷酸二酯寡核苷酸和正义RNA切割产物。释放的反义磷酸二酯寡脱氧核苷酸与靶RNA结合。环状哑铃状RNA/DNA嵌合寡核苷酸比线性反义硫代磷酸寡核苷酸（S-ODN-gag-AUG）和缺口哑铃状RNA/DNA嵌合寡核苷酸具有更强的核酸酶抗性和细胞摄取。合成具有AUG起始密码子序列的CDRDON-gag -AUG，作为HIV-1 gag基因（779-801）的靶标，并使用PBMC测试抑制作用。与反义硫代磷酸寡核苷酸（S-ODN-gag-AUG）相比，环状哑铃状RNA/DNA嵌合寡核苷酸（CDRDON-gag-AUG）显示出高度抑制作用，表明对HIV-1复制的序列特异性抑制，而不抑制逆转录酶和/或病毒进入过程，如反义硫代磷酸寡核苷酸。

项目成果

K.Takahashi, S.Baba, P.Chattopadhyay, Y.Koyanagi, N.Yamamoto, H.Takaku, and G.Kawai: "Structural Requirement for the Two-Step Dimerization of Human Immunodeficiency Virus Type-1 Genome."RNA. (in press.).

K.Takahashi、S.Baba、P.Chattopadhyay、Y.Koyanagi、N.Yamamoto、H.Takaku 和 G.Kawai：“人类免疫缺陷病毒 1 型基因组两步二聚化的结构要求”RNA。

Y.Tamura, M.Tao, N.Miyano-Kurosaki, K.Takai and H. Takaku: "Inhibition of human telomerase activity by antisense phosphorothioate oligonucleotides encapsulated with the transfection reagent, FuGENEィイD1TMィエD16, in HeLa cells."Antisense and Nucleic Acid Dru

Y.Tamura、M.Tao、N.Miyano-Kurosaki、K.Takai 和 H. Takaku：“用转染试剂 FuGENE D1TM D16 封装的反义硫代磷酸寡核苷酸在 HeLa 细胞中抑制人端粒酶活性。”反义和核酸酸药”

H.Yamakawa, T.Abe, T.Saito, K.Takai, N.Yamamoto, and H.Takaku: "Properties of nicked and circular dumbbell RNA/DNA chimeric oligonucleotides containing antisense phos-phodiester oligodeoxynucleotides."Bioorganic & Medicinal Chem.. 6. 1025-1038 (1998)

H.Yamakawa、T.Abe、T.Saito、K.Takai、N.Yamamoto 和 H.Takaku：“含有反义磷酸二酯寡脱氧核苷酸的切口和环形哑铃 RNA/DNA 嵌合寡核苷酸的特性。”

T.Hatta, K.Takai, S.Nakada, T.Yokota, and H.Takaku: "Specific inhibition of influenza virus RNA polymerase and nucleoprotein genes expression by liposomally endocapsulated antisense phosphorothioate oligonucleotides:penetration and localization of oligonu

T.Hatta、K.Takai、S.Nakada、T.Yokota 和 H.Takaku：“脂质体内包反义硫代磷酸寡核苷酸对流感病毒 RNA 聚合酶和核蛋白基因表达的特异性抑制：寡核苷酸的渗透和定位

Y.Koyanagi, Y.Tanaka, J.Kira, M.Ito, K.Hioki, N.Misawa, Y.Kawano, K.Yamasaki, R.Tanaka, Y.Suzuki, Y.Ueyama, E.Terada, T.Tanaka, M.Miyasaka, T.Kobayashi, Y.Kumazawa, and N.Yamamoto: "Primary viremia and CNS inavasion with HIV-1 in a novel hu-PBL-immunodcfi

Y.小柳、Y.田中、J.Kira、M.Ito、K.Hioki、N.Misawa、Y.Kawano、K.Yamasaki、R.Tanaka、Y.Suzuki、Y.Ueyama、E.Terada、T.