Antisense Activities of Ribozymes in in vivo

体内核酶的反义活性

• 批准号: 03404061
• 负责人: OHTSUKA Eiko
• 金额: $ 14.78万
• 依托单位: Faculty of Pharmaceutical Sciences Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03404061/
• 关键词: RNA enzyme; Bio-catalysis; ras gene; cell transformation; NIH3T3 cell; 合成遺伝子; NIH3T3細胞; RNA切断; トランスフォ-メ-ション

We have designed hammerhead ribozymes that cleaved the c-Ha-ras mRNA activated at codon 12 (GGU*GUU).Plasmids containing the ribozyme genes were expressed under the control of long terminal repeats of Rous sarcoma virus in NIH3T3 cells transfected with the activated c-Ha-ras gene. These ribozymes were found to inhibit the foci formation (about 50%) by cleavage activities with the ribozyme rather than hybridization to the c-Ha-ras gene product. Further, when the activated c-Ha-ras gene was cotransfected with the ribozyme gene, expression of the activated gene was suppressed by ribozymes in morphologically flat cells.

我们设计了能切割c-Ha-ras基因第12位密码子激活的mRNA的锤头状核酶（GGU*GUU），并在转染c-Ha-ras基因的NIH 3 T3细胞中，在Rous肉瘤病毒长末端重复序列的控制下表达了含有核酶基因的质粒。发现这些核酶通过与核酶的切割活性而不是与c-Ha-ras基因产物的杂交来抑制焦点形成（约50%）。此外，当激活的c-Ha-ras基因与核酶基因共转染时，在形态扁平的细胞中，激活基因的表达被核酶抑制。

项目成果

K.Kido: "Sequence-Dependent Cleavage of DNA by Alkylation with Antisense Oligodeoxyribonucleotides Containing a 2-(N-Iodoacetylaminoethyl)Thio-Adenine." Nucleic Acids Res.20. 1339-1344 (1992)

K.Kido：“通过使用含有 2-(N-碘乙酰氨基乙基)硫代腺嘌呤的反义寡脱氧核糖核苷酸进行烷基化，实现 DNA 的序列依赖性切割。”

K.Kido: "Sequence-Dependent Cleavage of DNA by Alkylation with Antisense Oligodeoxyribonucleotides Containing a 2-(N-Iodoacetylaminoethyl)Thio-Aenine." Nucleic Acids Res.20. 1339-1344 (1992)

K.Kido：“通过用含有 2-(N-碘乙酰氨基乙基)硫代-Aenine 的反义寡脱氧核糖核苷酸进行烷基化，实现 DNA 的序列依赖性切割。”

Hiroyuki Kamiya: "Transformation Activity of a Synthetic cーHaーras Gene Containing O^6ーMethylguanine in Codon 12" Jpn.J.Cancer Res.82. 997-1002 (1991)

Hiroyuki Kamiya：“密码子 12 中含有 O^6ー甲基鸟嘌呤的合成 cHaras 基因的转化活性”Jpn.J.Cancer Res.82 (1991)。

M.Koizumi: "Effects of Phosphorothioate and 2-Amino Groups in Hammerhead Ribozymes on Cleavage Rates and Mg^<2+>Binding." Biochemistry. 30. 5145-5150 (1991)

M.Koizumi：“锤头核酶中硫代磷酸酯和 2-氨基对裂解率和 Mg^<2> 结合的影响。”

M.Koezumi: "Effects of Plosphorothioate and 2-Amino Groups in Hammerhead Ribozymes on Cleavage Rates and Mg^<2+>Binding." Biochemistry. 30. 5145-5150 (1991)

M.Koezumi：“锤头核酶中硫代磷酸酯和 2-氨基对裂解率和 Mg^<2> 结合的影响。”