Formation of a catalytically active dimer by tRNAVal-driven short ribozymes

被引：62

作者：

Kuwabara, T

Warashina, M

Orita, M

Koseki, S

Ohkawa, J

Taira, K ^{[1
]}

机构：

[1] Natl Inst Adv Interdisciplinary Res, Tsukuba Sci City 3058562, Japan

[2] MITI, Agcy Ind Sci & Technol, Natl Inst Biosci & Human Technol, Tsukuba Sci City 3058566, Japan

[3] Yamanouchi Pharmaceut Co Ltd, Tsukuba Sci City 3050841, Japan

[4] Univ Tsukuba, Inst Appl Biochem, Tsukuba Sci City 3058572, Japan

来源：

NATURE BIOTECHNOLOGY | 1998年 / 16卷 / 10期

关键词：

antisense; minizymes; gene-inactivation;

D O I：

10.1038/nbt1098-961

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A minizyme is a hammerhead ribozyme with a short oligonucleotide linker instead of stem/loop II. Minizymes with low activity as monomers form active dimeric structures with a common stem. We explored the use of dimeric minizymes as gene-inactivating agents by placing minizymes under the control of a tRNA(Val) promoter. The tRNA(Val) portion of the transcript did not hinder dimerization as the tRNA-embedded minizyme formed an active dimeric structure. The cleavage activity of this minizyme that had been expressed either in vitro or in HeLa cells was almost one order of magnitude higher than that of the tRNA(Val)-embedded conventional hammerhead ribozyme. The tRNA(Val)-driven minizyme inhibited reporter gene activity (95%) whereas the tRNA(Val)-driven hammerhead ribozyme resulted in approximately 55% inhibition.

引用

页码：961 / 965

页数：5

共 42 条

[1] RNA ENZYME-DIRECTED GENE-THERAPY [J].