Sequence-specific cleavage of RNA using chimeric DNA splints and RNase H.
Identifieur interne : 002A70 ( PubMed/Corpus ); précédent : 002A69; suivant : 002A71Sequence-specific cleavage of RNA using chimeric DNA splints and RNase H.
Auteurs : H. Inoue ; Y. Hayase ; S. Iwai ; E. OhtsukaSource :
- Nucleic acids symposium series [ 0261-3166 ] ; 1988.
English descriptors
- KwdEn :
- MESH :
- chemical : DNA, Endoribonucleases, Oligodeoxyribonucleotides, RNA, Ribonuclease H.
- enzymology : Escherichia coli.
- Base Sequence, Chimera, Molecular Sequence Data, Substrate Specificity.
Abstract
To cleave RNA molecules using E. coli RNase H in a site-specific manner, a short oligodeoxyribonucleotide (3-5 mer) linked with oligo(2'-O-methyl)ribonucleotide(s) was designed to be used as a DNA splint. Our model experiments with ribooligomer the splint duplexes (9 mers) and RNase H demonstrated that a tetradeoxynucleotide cluster seems to be sufficient for the enzyme recognition and the short DNA-containing splint directs a unique cleavage of RNA by RNase H. The method could be applied to longer ribooligonucleotide substrates. For example, when 3'm (GA)d(AGAA)m(GGU)5' was used as a hybridization strand, 32pUCUUUCUUCUUCCAGGAU was cleaved specifically between U11 and C12 to yield 32pUCUUUCUUCUU. This method will have a variety of applications for the study of RNA.
PubMed: 2465538
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pubmed:2465538Le document en format XML
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<author><name sortKey="Inoue, H" sort="Inoue, H" uniqKey="Inoue H" first="H" last="Inoue">H. Inoue</name>
<affiliation><nlm:affiliation>Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.</nlm:affiliation>
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<author><name sortKey="Hayase, Y" sort="Hayase, Y" uniqKey="Hayase Y" first="Y" last="Hayase">Y. Hayase</name>
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<author><name sortKey="Iwai, S" sort="Iwai, S" uniqKey="Iwai S" first="S" last="Iwai">S. Iwai</name>
</author>
<author><name sortKey="Ohtsuka, E" sort="Ohtsuka, E" uniqKey="Ohtsuka E" first="E" last="Ohtsuka">E. Ohtsuka</name>
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<author><name sortKey="Hayase, Y" sort="Hayase, Y" uniqKey="Hayase Y" first="Y" last="Hayase">Y. Hayase</name>
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<author><name sortKey="Iwai, S" sort="Iwai, S" uniqKey="Iwai S" first="S" last="Iwai">S. Iwai</name>
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<author><name sortKey="Ohtsuka, E" sort="Ohtsuka, E" uniqKey="Ohtsuka E" first="E" last="Ohtsuka">E. Ohtsuka</name>
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<series><title level="j">Nucleic acids symposium series</title>
<idno type="ISSN">0261-3166</idno>
<imprint><date when="1988" type="published">1988</date>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Base Sequence</term>
<term>Chimera</term>
<term>DNA</term>
<term>Endoribonucleases</term>
<term>Escherichia coli (enzymology)</term>
<term>Molecular Sequence Data</term>
<term>Oligodeoxyribonucleotides</term>
<term>RNA</term>
<term>Ribonuclease H</term>
<term>Substrate Specificity</term>
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<term>Endoribonucleases</term>
<term>Oligodeoxyribonucleotides</term>
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<term>Ribonuclease H</term>
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<keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Escherichia coli</term>
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<keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term>
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<front><div type="abstract" xml:lang="en">To cleave RNA molecules using E. coli RNase H in a site-specific manner, a short oligodeoxyribonucleotide (3-5 mer) linked with oligo(2'-O-methyl)ribonucleotide(s) was designed to be used as a DNA splint. Our model experiments with ribooligomer the splint duplexes (9 mers) and RNase H demonstrated that a tetradeoxynucleotide cluster seems to be sufficient for the enzyme recognition and the short DNA-containing splint directs a unique cleavage of RNA by RNase H. The method could be applied to longer ribooligonucleotide substrates. For example, when 3'm (GA)d(AGAA)m(GGU)5' was used as a hybridization strand, 32pUCUUUCUUCUUCCAGGAU was cleaved specifically between U11 and C12 to yield 32pUCUUUCUUCUU. This method will have a variety of applications for the study of RNA.</div>
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<DateCompleted><Year>1989</Year>
<Month>03</Month>
<Day>29</Day>
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<DateRevised><Year>2007</Year>
<Month>11</Month>
<Day>15</Day>
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<Title>Nucleic acids symposium series</Title>
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<ArticleTitle>Sequence-specific cleavage of RNA using chimeric DNA splints and RNase H.</ArticleTitle>
<Pagination><MedlinePgn>135-8</MedlinePgn>
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<Abstract><AbstractText>To cleave RNA molecules using E. coli RNase H in a site-specific manner, a short oligodeoxyribonucleotide (3-5 mer) linked with oligo(2'-O-methyl)ribonucleotide(s) was designed to be used as a DNA splint. Our model experiments with ribooligomer the splint duplexes (9 mers) and RNase H demonstrated that a tetradeoxynucleotide cluster seems to be sufficient for the enzyme recognition and the short DNA-containing splint directs a unique cleavage of RNA by RNase H. The method could be applied to longer ribooligonucleotide substrates. For example, when 3'm (GA)d(AGAA)m(GGU)5' was used as a hybridization strand, 32pUCUUUCUUCUUCCAGGAU was cleaved specifically between U11 and C12 to yield 32pUCUUUCUUCUU. This method will have a variety of applications for the study of RNA.</AbstractText>
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