Fine-tuning of ENA gapmers as antisense oligonucleotides for sequence-specific inhibition.

Fine-tuning of ENA gapmers as antisense oligonucleotides for sequence-specific inhibition. Research Abstract Details 

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Fine-tuning of ENA gapmers as antisense oligonucleotides for sequence-specific inhibition. Abstract Text:

Miho Takagi-Sato,Shinya Tokuhiro,Reimi Kawaida,Makoto Koizumi,

For gene validation and the development of oligonucleotide agents, 2'-O,4'-C-ethylene-bridged nucleic acid (ENA) antisense gapmers are widely available. An in vitro Escherichia coli RNase H reaction analysis using ENA gapmers and an RNA oligonucleotide with mouse peptidylarginine deiminase 4 (PADI4) gene sequences revealed that the RNA oligonucleotide was specifically cleaved in the only reported case of the use of an ENA gapmer with an antisense sequence. On the other hand, duplexes of the full-length transcripts of PADI4 mRNA and ENA gapmers with a wide DNA window were cleaved not only at the target site, but also at nontarget sites by RNase H derived from partial base-pairing between the transcript and the ENA gapmer. When the DNA window region of the ENA gapmer was shortened to 5 or 6 nucleotides, the nontarget cleavage was effectively diminished. Moreover, the specific inhibition of PADI4 mRNA expression was observed in the cotransfection of PADI4 cDNA and ENA gapmers containing a short DNA region into NIH3T3 cells. These results demonstrated that ENA gapmers with a short DNA region improved the sequence-specificity of mRNA downregulation. These optimized ENA gapmers could reduce the "off-target" effect and be applicable to gene validation and oligonucleotide therapeutics.

Fine-tuning of ENA gapmers as antisense oligonucleotides for sequence-specific inhibition. Publishing Authors By Initials

M Takagi-Sato,S Tokuhiro,R Kawaida,M Koizumi,

For similar enzymes and coenzymes: enzymes: hydrolases: esterases: endonucleases: endoribonucleases: ribonuclease h research abstracts see: enzymes and coenzymes: enzymes: hydrolases: esterases: endonucleases: endoribonucleases: ribonuclease h research

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Fine-tuning of ENA gapmers as antisense oligonucleotides for sequence-specific inhibition. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Oligonucleotides

VOLUME: 17

Page Numbers: 291-301

Journal Abbreviation:

ISSN: 1545-4576

DAY: 26

MONTH: 11

YEAR: 2007

Fine-tuning of ENA gapmers as antisense oligonucleotides for sequence-specific inhibition. Information

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LANGUAGE: eng

NlmUniqueID: 101188415

Fine-tuning of ENA gapmers as antisense oligonucleotides for sequence-specific inhibition. Keywords Mesh Terms:

KEYWORDS: Ribonuclease H

MESH TERMS: metabolism

Chemical & Substance for Abstract: Fine-tuning of ENA gapmers as antisense oligonucleotides for sequence-specific inhibition. Information

Substance Name: peptidylarginine deiminase type IV

Registry Number: EC 3.5.3.15

Grant and Affiliation Information for Fine-tuning of ENA gapmers as antisense oligonucleotides for sequence-specific inhibition.

AFFILIATION: Medicinal Chemistry Research Laboratories I, Daiichi Sankyo Co., Ltd., 1-2-58, Hiromachi, Shinagawa-ku, Tokyo 140-8710, Japan.

Country: United States

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MEDLINETA: Oligonucleotides

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