FUSION WITH AN RNA BINDING DOMAIN TO CONFER TARGET RNA SPECIFICITY TO AN RNASE - DESIGN AND ENGINEERING OF TAT-RNASE H THAT SPECIFICALLY RECOGNIZES AND CLEAVES HIV-1 RNA IN VITRO

Melekhovets YF; Joshi S

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FUSION WITH AN RNA BINDING DOMAIN TO CONFER TARGET RNA SPECIFICITY TO AN RNASE - DESIGN AND ENGINEERING OF TAT-RNASE H THAT SPECIFICALLY RECOGNIZES AND CLEAVES HIV-1 RNA IN VITRO

机译：与RNA结合域融合以赋予RNASE特定的目标RNA特异性-TAT-RNASE H的设计和工程，可特异性地识别并清除HIV-1 RNA

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A target RNA/DNA-specific nuclease could be constructed if a specific RNA/DNA binding domain allowing target RNA/DNA recognition was fused to a (deoxy)ribonucleolytic domain allowing target RNA/DNA cleavage. The design and construction of such a chimeric enzyme could be of value for both basic research involving structure-function relationships and applied research requiring inactivation of harmful RNA/DNA molecules of cellular or pathogenic origin. The feasibility of this designer nuclease approach for inactivating specific RNA/DNA molecules was assessed using human immunodeficiency virus type-1 (HIV-1) RNA as a model. Trans-activator of transcription (Tat) protein is one of the key regulatory proteins encoded by HIV-1. It binds to the trans-activation-responsive (TAR) RNA element located within the 5' non-coding region of HIV-1 RNAs. The TAR RNA binding domain of this protein was fused to the ribonuclease (RNase) H domain of HIV-1 reverse transcriptase (RT). RNase H by itself lacks an RNA binding domain. The chimeric Tat-RNase H protein was shown to specifically recognize and cleave HIV-1 TAR RNA in vitro. Cleavage was abolished by mutations in the Tat binding region within the TAR RNA, indicating that it is specific to HIV-1 TAR RNA.

机译：如果允许靶RNA / DNA识别的特定RNA / DNA结合域与允许靶RNA / DNA裂解的（脱氧）核糖核糖核酸融合域融合，则可以构建靶RNA / DNA特异性核酸酶。这种嵌合酶的设计和构建对于涉及结构-功能关系的基础研究和要求灭活细胞或致病性有害RNA / DNA分子的应用研究都可能具有价值。使用人类免疫缺陷病毒1型（HIV-1）RNA作为模型，评估了该设计者核酸酶方法灭活特定RNA / DNA分子的可行性。转录反式激活蛋白（Tat）是HIV-1编码的关键调节蛋白之一。它与位于HIV-1 RNA 5'非编码区内的反式激活（TAR）RNA元件结合。该蛋白的TAR RNA结合结构域与HIV-1逆转录酶（RT）的核糖核酸酶（RNase）H结构域融合。 RNase H本身缺乏RNA结合域。嵌合的Tat-RNase H蛋白显示出在体外特异性识别和切割HIV-1 TAR RNA的能力。 TAR RNA内Tat结合区的突变消除了切割，表明它对HIV-1 TAR RNA具有特异性。

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《Nucleic Acids Research》 |1996年第10期|共5页
作者
Melekhovets YF; Joshi S;
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正文语种 eng
中图分类细胞形态学;生物化学;
关键词
Human immunodeficiency virus; Reverse transcriptase; Mutational; analysis; Type 1; Dna; Transactivation; Purification; Inhibition; Expression; Polymerase;

机译：人类免疫缺陷病毒;逆转录酶;突变;分析;1型;Dna;反式激活;纯化;抑制;表达;聚合酶;

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专利

1. FUSION WITH AN RNA BINDING DOMAIN TO CONFER TARGET RNA SPECIFICITY TO AN RNASE - DESIGN AND ENGINEERING OF TAT-RNASE H THAT SPECIFICALLY RECOGNIZES AND CLEAVES HIV-1 RNA IN VITRO [J] . Melekhovets YF, Joshi S Nucleic Acids Research . 1996,第10期

机译：与RNA结合域融合以赋予RNASE特定的目标RNA特异性-TAT-RNASE H的设计和工程，可特异性地识别并清除HIV-1 RNA
2. Fusion With an RNA Binding Domain to Confer Target RNA Specificity to an RNase: Design and Engineering of Tat-RNase H That Specifically Recognizes and Cleaves HIV-1 RNA in vitro [J] . Sadhna Joshi, Yuri F. Melekhovets Nucleic acids research . 1996,第10期

机译：与RNA结合域融合，赋予RNase靶RNA特异性：Tat-RNase H的设计和工程，该Tat-RNase H在体外特异性识别和裂解HIV-1 RNA。
3. Fusion With an RNA Binding Domain to Confer Target RNA Specificity to an RNase: Design and Engineering of Tat-RNase H That Specifically Recognizes and Cleaves HIV-1 RNA in vitro [J] . Sadhna Joshi, Yuri F. Melekhovets Nucleic acids research . 1996,第10期

机译：与RNA结合域融合，赋予RNase靶RNA特异性：Tat-RNase H的设计和工程，该Tat-RNase H在体外特异性识别和裂解HIV-1 RNA。
4. Design, Synthesis and RNase H activity of Hemi-Gapmer Type Peptide Ribonucleic Acid (PRNA)-DNA chimeras for the RNase H mediated catalytic oligonucleotide therapeutics [C] . Masahito Inagaki, Daisuke Unabara, Ryohei Uematsu 日本化学会春季年会講演予稿集 . 2018

机译：半介距型肽核糖核糖核酸（PRNA）-DNA嵌合催化寡核苷酸治疗剂的设计，合成和RNase H活性
5. Investigating the RNA binding domains of MBNL1 and the alternative splicing motifs they recognize [D] . Purcell, Jamie 2012

机译：研究MBNL1的RNA结合结构域和它们识别的替代剪接基序
6. Fusion with an RNA binding domain to confer target RNA specificity to an RNase: design and engineering of Tat-RNase H that specifically recognizes and cleaves HIV-1 RNA in vitro. [O] . Y F Melekhovets, S Joshi 1996

机译：与RNA结合域融合以赋予RNA酶以靶RNA特异性：Tat-RNase H的设计和工程设计可以在体外特异性识别和切割HIV-1 RNA。
7. Fusion with an RNA binding domain to confer target RNA specificity to an RNase: design and engineering of Tat-RNase H that specifically recognizes and cleaves HIV-1 RNA in vitro. [O] . Melekhovets, Y F, Joshi, S 1996

机译：与RNA结合域融合以赋予RNA酶以靶RNA特异性：Tat-RNase H的设计和工程设计，可以在体外特异性识别和切割HIV-1 RNA。

FUSION WITH AN RNA BINDING DOMAIN TO CONFER TARGET RNA SPECIFICITY TO AN RNASE - DESIGN AND ENGINEERING OF TAT-RNASE H THAT SPECIFICALLY RECOGNIZES AND CLEAVES HIV-1 RNA IN VITRO

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