Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.26.4 (RNase H)
 2,751 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In order to evaluate the function of the mouse peptidylarginine deiminase 4 (PADI4) gene, gapmer-designed ENA antisense oligonucleotides were utilized. Antisense ENA gapmers were found to inhibit the mRNA expression of the PADI4 gene, but ENA gapmers with sense sequences as controls only partially inhibited this mRNA expression. An in vitro E. coli RNase H reaction analysis using transcripts of the PADI4 gene and their ENA gapmers revealed that the mixtures of the transcripts and ENA gapmers with sense sequences were cleaved at non-target sites by RNase H derived from partial base-pairing between the transcript and the ENA gapmer. In an in vitro E. coli RNase H reaction analysis, when the DNA region of the ENA gapmer was shortened to 5 or 6 nucleotides, the non-target cleavage disappeared and the specific inhibition of PADI4 mRNA expression was observed. These results demonstrated that ENA gapmers with a short DNA region improved the sequence-specificity of mRNA down-regulation. As well, they suggest that we should be alert to the utility of antisense oligonucleotides with a wide DNA region in terms of sequence specificity, and additionally, that optimized ENA gapmers could be useful for application to gene validation.
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PMID:Design of ENA gapmers as fine-tuning antisense oligonucleotides with sequence-specific inhibitory activity on mouse PADI4 mRNA expression. 1715 Sep 46

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.
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PMID:Fine-tuning of ENA gapmers as antisense oligonucleotides for sequence-specific inhibition. 1785 69