Specific inhibition of in vitro reverse transcription using antisense oligonucleotides targeted to the TAR regions of HIV-1 and HIV-2
Identifieur interne : 000913 ( Istex/Corpus ); précédent : 000912; suivant : 000914Specific inhibition of in vitro reverse transcription using antisense oligonucleotides targeted to the TAR regions of HIV-1 and HIV-2
Auteurs : Florence Boulme ; Maritta Per L Heape ; Leila Sarih-Cottin ; Simon LitvakSource :
- BBA - Gene Structure and Expression [ 0167-4781 ] ; 1997.
English descriptors
- Teeft :
- Acta, Antisense, Antisense odns, Antisense oligonucleotides, Biochimica, Biol, Biophysica, Biophysica acta, Blockage, Boulme, Cdna, Cdna synthesis, Control odns, Important role, Nucleic acids, Odns, Oligonucleotides, Primer, Replication, Retroviral, Rnase, Secondary structure, Target sequence, Transcription, Viral.
Abstract
Abstract: Antisense oligonucleotides (ODNs) overlapping the stem-loop structure of the trans-activating responsive (TAR) element at the 5′ end of HIV-1 and HIV-2 viral RNAs were tested for their inhibitory effect on cDNA synthesis by HIV-1 and HIV-2 reverse transcriptases (RT). Inhibition of reverse transcription is sequence-specific and enhanced by the presence of the RT-associated RNase H activity. The degree of inhibition obtained with the anti-TAR antisense is significantly higher than with other HIV-1 targeted antisense ODNs used before [1]. Gel retardation showed a stable specific complex between the 16- and 25-mer anti-TAR HIV-1 selected ODNs and the target region. No complex was observed with a non-inhibitor 22-mer anti-TAR ODN and with the corresponding control sequences. Targeting of the first stem-loop in the 5′ region of HIV-2 RNA by anti-TAR ODNs inhibited very strongly reverse transcription by HIV-2 RT. The structure of the antisense and the target sequence affect annealing efficiency and hence the degree of inhibition of reverse transcription. © 1997 Elsevier Science B.V. All rights reserved.
Url:
DOI: 10.1016/S0167-4781(97)00026-2
Links to Exploration step
ISTEX:191164A3CCA1C0429CDAB785CEE915EA596FE97ELe document en format XML
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Gene Structure and Expression</title><title level="j" type="abbrev">BBAEXP</title><idno type="ISSN">0167-4781</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1997">1997</date><biblScope unit="volume">1351</biblScope><biblScope unit="issue">3</biblScope><biblScope unit="page" from="249">249</biblScope><biblScope unit="page" to="255">255</biblScope></imprint><idno type="ISSN">0167-4781</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0167-4781</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Acta</term><term>Antisense</term><term>Antisense odns</term><term>Antisense oligonucleotides</term><term>Biochimica</term><term>Biol</term><term>Biophysica</term><term>Biophysica acta</term><term>Blockage</term><term>Boulme</term><term>Cdna</term><term>Cdna synthesis</term><term>Control odns</term><term>Important role</term><term>Nucleic acids</term><term>Odns</term><term>Oligonucleotides</term><term>Primer</term><term>Replication</term><term>Retroviral</term><term>Rnase</term><term>Secondary structure</term><term>Target sequence</term><term>Transcription</term><term>Viral</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Antisense oligonucleotides (ODNs) overlapping the stem-loop structure of the trans-activating responsive (TAR) element at the 5′ end of HIV-1 and HIV-2 viral RNAs were tested for their inhibitory effect on cDNA synthesis by HIV-1 and HIV-2 reverse transcriptases (RT). Inhibition of reverse transcription is sequence-specific and enhanced by the presence of the RT-associated RNase H activity. The degree of inhibition obtained with the anti-TAR antisense is significantly higher than with other HIV-1 targeted antisense ODNs used before [1]. Gel retardation showed a stable specific complex between the 16- and 25-mer anti-TAR HIV-1 selected ODNs and the target region. No complex was observed with a non-inhibitor 22-mer anti-TAR ODN and with the corresponding control sequences. Targeting of the first stem-loop in the 5′ region of HIV-2 RNA by anti-TAR ODNs inhibited very strongly reverse transcription by HIV-2 RT. The structure of the antisense and the target sequence affect annealing efficiency and hence the degree of inhibition of reverse transcription. © 1997 Elsevier Science B.V. All rights reserved.</div></front></TEI><istex><corpusName>elsevier</corpusName><keywords><teeft><json:string>antisense</json:string><json:string>odns</json:string><json:string>rnase</json:string><json:string>cdna</json:string><json:string>antisense odns</json:string><json:string>cdna synthesis</json:string><json:string>primer</json:string><json:string>retroviral</json:string><json:string>oligonucleotides</json:string><json:string>viral</json:string><json:string>biol</json:string><json:string>biophysica acta</json:string><json:string>biophysica</json:string><json:string>blockage</json:string><json:string>boulme</json:string><json:string>biochimica</json:string><json:string>acta</json:string><json:string>transcription</json:string><json:string>important role</json:string><json:string>target sequence</json:string><json:string>secondary structure</json:string><json:string>replication</json:string><json:string>antisense oligonucleotides</json:string><json:string>control odns</json:string><json:string>nucleic acids</json:string></teeft></keywords><author><json:item><name>Florence Boulme</name><affiliations><json:string>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</json:string></affiliations></json:item><json:item><name>Maritta Perälä-Heape</name><affiliations><json:string>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</json:string></affiliations></json:item><json:item><name>Leila Sarih-Cottin</name><affiliations><json:string>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</json:string></affiliations></json:item><json:item><name>Simon Litvak</name><affiliations><json:string>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>HIV-1/HIV-2</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Replication</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Reverse transcription</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Inhibition</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Antisense</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Trans-activating region</value></json:item></subject><arkIstex>ark:/67375/6H6-06V08SB7-Z</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>Full-length article</json:string></originalGenre><abstract>Abstract: Antisense oligonucleotides (ODNs) overlapping the stem-loop structure of the trans-activating responsive (TAR) element at the 5′ end of HIV-1 and HIV-2 viral RNAs were tested for their inhibitory effect on cDNA synthesis by HIV-1 and HIV-2 reverse transcriptases (RT). Inhibition of reverse transcription is sequence-specific and enhanced by the presence of the RT-associated RNase H activity. The degree of inhibition obtained with the anti-TAR antisense is significantly higher than with other HIV-1 targeted antisense ODNs used before [1]. Gel retardation showed a stable specific complex between the 16- and 25-mer anti-TAR HIV-1 selected ODNs and the target region. No complex was observed with a non-inhibitor 22-mer anti-TAR ODN and with the corresponding control sequences. Targeting of the first stem-loop in the 5′ region of HIV-2 RNA by anti-TAR ODNs inhibited very strongly reverse transcription by HIV-2 RT. The structure of the antisense and the target sequence affect annealing efficiency and hence the degree of inhibition of reverse transcription. © 1997 Elsevier Science B.V. All rights reserved.</abstract><qualityIndicators><score>7.729</score><pdfWordCount>3725</pdfWordCount><pdfCharCount>21837</pdfCharCount><pdfVersion>1.1</pdfVersion><pdfPageCount>7</pdfPageCount><pdfPageSize>595 x 763 pts</pdfPageSize><refBibsNative>true</refBibsNative><abstractWordCount>167</abstractWordCount><abstractCharCount>1124</abstractCharCount><keywordCount>6</keywordCount></qualityIndicators><title>Specific inhibition of in vitro reverse transcription using antisense oligonucleotides targeted to the TAR regions of HIV-1 and HIV-2</title><pmid><json:string>9130587</json:string></pmid><pii><json:string>S0167-4781(97)00026-2</json:string></pii><genre><json:string>research-article</json:string></genre><host><title>BBA - Gene Structure and Expression</title><language><json:string>unknown</json:string></language><publicationDate>1997</publicationDate><issn><json:string>0167-4781</json:string></issn><pii><json:string>S0167-4781(00)X0033-4</json:string></pii><volume>1351</volume><issue>3</issue><pages><first>249</first><last>255</last></pages><genre><json:string>journal</json:string></genre></host><namedEntities><unitex><date><json:string>1997</json:string></date><geogName></geogName><orgName><json:string>Ministry of Education and Research</json:string><json:string>F</json:string><json:string>SIDACTION</json:string><json:string>Science Academy of Finland</json:string><json:string>Pole Medicament</json:string><json:string>ANRS</json:string></orgName><orgName_funder><json:string>F</json:string><json:string>SIDACTION</json:string><json:string>Science Academy of Finland</json:string><json:string>Pole Medicament</json:string><json:string>ANRS</json:string></orgName_funder><orgName_provider></orgName_provider><persName><json:string>Leila Sarih-Cottin</json:string><json:string>J. Toulme</json:string><json:string>Christina Calmels</json:string><json:string>Jean Luc</json:string><json:string>Daniele Tharaud</json:string><json:string>Florence Boulme</json:string><json:string>Roger Goody</json:string><json:string>Jean-Luc Darlix</json:string><json:string>Žfor</json:string><json:string>Camille Saint-Saens</json:string><json:string>Max Planck</json:string><json:string>Žresults</json:string><json:string>Simon Litvak</json:string></persName><placeName><json:string>Dortmund</json:string><json:string>Germany</json:string><json:string>Finland</json:string><json:string>Oulu</json:string><json:string>France</json:string><json:string>Lyon</json:string></placeName><ref_url></ref_url><ref_bibl><json:string>F. Boulme et al.</json:string><json:string>Vickers et al.</json:string></ref_bibl><bibl></bibl></unitex></namedEntities><ark><json:string>ark:/67375/6H6-06V08SB7-Z</json:string></ark><categories><wos></wos><scienceMetrix><json:string>1 - health sciences</json:string><json:string>2 - biomedical research</json:string><json:string>3 - biochemistry & molecular biology</json:string></scienceMetrix><scopus><json:string>1 - Life Sciences</json:string><json:string>2 - Biochemistry, Genetics and Molecular Biology</json:string><json:string>3 - Genetics</json:string><json:string>1 - Life Sciences</json:string><json:string>2 - Biochemistry, Genetics and Molecular Biology</json:string><json:string>3 - Biochemistry</json:string><json:string>1 - Life Sciences</json:string><json:string>2 - Biochemistry, Genetics and Molecular Biology</json:string><json:string>3 - Biophysics</json:string><json:string>1 - Life Sciences</json:string><json:string>2 - Biochemistry, Genetics and Molecular Biology</json:string><json:string>3 - Structural Biology</json:string></scopus><inist><json:string>1 - sciences appliquees, technologies et medecines</json:string><json:string>2 - sciences biologiques et medicales</json:string><json:string>3 - sciences medicales</json:string></inist></categories><publicationDate>1997</publicationDate><copyrightDate>1997</copyrightDate><doi><json:string>10.1016/S0167-4781(97)00026-2</json:string></doi><id>191164A3CCA1C0429CDAB785CEE915EA596FE97E</id><score>1</score><fulltext><json:item><extension>pdf</extension><original>true</original><mimetype>application/pdf</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-06V08SB7-Z/fulltext.pdf</uri></json:item><json:item><extension>zip</extension><original>false</original><mimetype>application/zip</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-06V08SB7-Z/bundle.zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/ark:/67375/6H6-06V08SB7-Z/fulltext.tei"><teiHeader><fileDesc><titleStmt><title level="a">Specific inhibition of in vitro reverse transcription using antisense oligonucleotides targeted to the TAR regions of HIV-1 and HIV-2</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher scheme="https://scientific-publisher.data.istex.fr">ELSEVIER</publisher><availability><licence><p>©1997 Elsevier Science B.V.</p></licence><p scheme="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</p></availability><date>1997</date></publicationStmt><notesStmt><note type="research-article" scheme="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</note><note type="journal" scheme="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</note><note type="content">Section title: Rapid report</note><note type="content">Fig. 1: A: Antisense oligonucleotides (25-, 22- and 16-mers) directed against the HIV-1 TAR element are shown covering different regions of the stem-loop of the TAR RNA secondary structure. B: Schematic representation of the expected sizes of the HIV-1 cDNA products synthesized by HIV-1 RT from an anti-PBS primer in the presence of anti-TAR oligonucleotides of different lengths.</note><note type="content">Fig. 2: Effect of HIV-1 anti-TAR antisense ODNs on the initiation of reverse transcription. cDNA synthesis was performed either by HIV-RT (RNase H−), or (RNase H+) expressed and purified as described before [16–18]. M is the lane corresponding to the size markers in nucleotides. A: cDNA synthesis performed by HIV-1 RT (RNase H−), in the presence of anti-PBS 18-mer primer. Lane 1, control (cDNA synthesis in the presence of anti-PBS); lanes 2 and 5, anti-TAR 25-mer, respectively non-blocked (n) or blocked (b) in its 3′ end; lanes 3 and 6, anti-TAR 22-mer n and b; lanes 4 and 7, anti-TAR 16-mer n and b; antisense ODNs are used at a concentration of 1 μM. B: cDNA synthesis by HIV-RT p66/p51 (RNase H+) in the presence of anti-PBS 18-mer primer. Lane 1, control; lanes 2, 3 and 4, anti-TAR 25-mer at 0.5, 1.0 and 5.0 μM; lane 5, anti-TAR 22-mer at 1 μM; lanes 6, 7 and 8, anti-TAR 16-mer at 0.5, 1.0 and 5.0 μM, respectively. C: Effect of different concentrations of anti-TAR 25-mer antisense ODNs on the initiation of reverse transcription performed by HIV-RT (RNase H−) (□), or p66/p51 (RNase H+) (■).</note><note type="content">Fig. 3: Gel mobility shift of antisense/RNA target duplexes. Conditions are described in the text. Lane C: control labelled RNA alone; primer: anti-PBS (18-mer); lanes 1 and 2: anti-TAR (16-mer); lanes 3 and 4: anti-TAR (22-mer); lanes 5 and 6: anti-TAR (25-mer); lanes 7 and 8: mismatched anti-TAR (25-mer); lanes 9 and 10: scrambled anti-TAR (25-mer). Lanes 1, 3, 5, 7 and 9: 0,5 μM; lanes 2, 4, 6, 8 and 10: 1 μM. Experimental details are described in the text.</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a">Specific inhibition of in vitro reverse transcription using antisense oligonucleotides targeted to the TAR regions of HIV-1 and HIV-2</title><author xml:id="author-0000"><persName><forename type="first">Florence</forename><surname>Boulme</surname></persName><affiliation>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</affiliation></author><author xml:id="author-0001"><persName><forename type="first">Maritta</forename><surname>Perälä-Heape</surname></persName><note type="biography">Present address: Fibrogen Europe, Kiviharjunke 11, 90220 Oulu, Finland.</note><affiliation>Present address: Fibrogen Europe, Kiviharjunke 11, 90220 Oulu, Finland.</affiliation><affiliation>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</affiliation></author><author xml:id="author-0002"><persName><forename type="first">Leila</forename><surname>Sarih-Cottin</surname></persName><affiliation>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</affiliation></author><author xml:id="author-0003"><persName><forename type="first">Simon</forename><surname>Litvak</surname></persName><affiliation>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</affiliation></author><idno type="istex">191164A3CCA1C0429CDAB785CEE915EA596FE97E</idno><idno type="ark">ark:/67375/6H6-06V08SB7-Z</idno><idno type="DOI">10.1016/S0167-4781(97)00026-2</idno><idno type="PII">S0167-4781(97)00026-2</idno></analytic><monogr><title level="j">BBA - Gene Structure and Expression</title><title level="j" type="abbrev">BBAEXP</title><idno type="pISSN">0167-4781</idno><idno type="PII">S0167-4781(00)X0033-4</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1997"></date><biblScope unit="volume">1351</biblScope><biblScope unit="issue">3</biblScope><biblScope unit="page" from="249">249</biblScope><biblScope unit="page" to="255">255</biblScope></imprint></monogr></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>1997</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>Abstract: Antisense oligonucleotides (ODNs) overlapping the stem-loop structure of the trans-activating responsive (TAR) element at the 5′ end of HIV-1 and HIV-2 viral RNAs were tested for their inhibitory effect on cDNA synthesis by HIV-1 and HIV-2 reverse transcriptases (RT). Inhibition of reverse transcription is sequence-specific and enhanced by the presence of the RT-associated RNase H activity. The degree of inhibition obtained with the anti-TAR antisense is significantly higher than with other HIV-1 targeted antisense ODNs used before [1]. Gel retardation showed a stable specific complex between the 16- and 25-mer anti-TAR HIV-1 selected ODNs and the target region. No complex was observed with a non-inhibitor 22-mer anti-TAR ODN and with the corresponding control sequences. Targeting of the first stem-loop in the 5′ region of HIV-2 RNA by anti-TAR ODNs inhibited very strongly reverse transcription by HIV-2 RT. The structure of the antisense and the target sequence affect annealing efficiency and hence the degree of inhibition of reverse transcription. © 1997 Elsevier Science B.V. All rights reserved.</p></abstract><textClass><keywords scheme="keyword"><list><head>Keywords</head><item><term>HIV-1/HIV-2</term></item><item><term>Replication</term></item><item><term>Reverse transcription</term></item><item><term>Inhibition</term></item><item><term>Antisense</term></item><item><term>Trans-activating region</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1997-01-28">Modified</change><change when="1997">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-06V08SB7-Z/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: ce:floats; body; tail"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"><istex:entity SYSTEM="gr1" NDATA="IMAGE" name="gr1"></istex:entity><istex:entity SYSTEM="gr2" NDATA="IMAGE" name="gr2"></istex:entity><istex:entity SYSTEM="gr3" NDATA="IMAGE" name="gr3"></istex:entity></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla"><item-info><jid>BBAEXP</jid><aid>90995</aid><ce:pii>S0167-4781(97)00026-2</ce:pii><ce:doi>10.1016/S0167-4781(97)00026-2</ce:doi><ce:copyright year="1997" type="full-transfer">Elsevier Science B.V.</ce:copyright></item-info><head><ce:dochead><ce:textfn>Rapid report</ce:textfn></ce:dochead><ce:title>Specific inhibition of in vitro reverse transcription using antisense oligonucleotides targeted to the TAR regions of HIV-1 and HIV-2</ce:title><ce:author-group><ce:author><ce:given-name>Florence</ce:given-name><ce:surname>Boulme</ce:surname></ce:author><ce:author><ce:given-name>Maritta</ce:given-name><ce:surname>Perälä-Heape</ce:surname><ce:cross-ref refid="FN1"><ce:sup>1</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Leila</ce:given-name><ce:surname>Sarih-Cottin</ce:surname></ce:author><ce:author><ce:given-name>Simon</ce:given-name><ce:surname>Litvak</ce:surname><ce:cross-ref refid="CORR1">*</ce:cross-ref></ce:author><ce:affiliation><ce:textfn>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</ce:textfn></ce:affiliation><ce:correspondence id="CORR1"><ce:label>*</ce:label><ce:text>Corresponding author. Fax: +33 556999057; E-mail: simon.litvak@ibgc.u-bordeaux2.fr</ce:text></ce:correspondence><ce:footnote id="FN1"><ce:label>1</ce:label><ce:note-para>Present address: Fibrogen Europe, Kiviharjunke 11, 90220 Oulu, Finland.</ce:note-para></ce:footnote></ce:author-group><ce:date-received day="6" month="1" year="1997"></ce:date-received><ce:date-revised day="28" month="1" year="1997"></ce:date-revised><ce:date-accepted day="31" month="1" year="1997"></ce:date-accepted><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>Antisense oligonucleotides (ODNs) overlapping the stem-loop structure of the trans-activating responsive (TAR) element at the 5′ end of HIV-1 and HIV-2 viral RNAs were tested for their inhibitory effect on cDNA synthesis by HIV-1 and HIV-2 reverse transcriptases (RT). Inhibition of reverse transcription is sequence-specific and enhanced by the presence of the RT-associated RNase H activity. The degree of inhibition obtained with the anti-TAR antisense is significantly higher than with other HIV-1 targeted antisense ODNs used before <ce:cross-ref refid="BIB1">[1]</ce:cross-ref>. Gel retardation showed a stable specific complex between the 16- and 25-mer anti-TAR HIV-1 selected ODNs and the target region. No complex was observed with a non-inhibitor 22-mer anti-TAR ODN and with the corresponding control sequences. Targeting of the first stem-loop in the 5′ region of HIV-2 RNA by anti-TAR ODNs inhibited very strongly reverse transcription by HIV-2 RT. The structure of the antisense and the target sequence affect annealing efficiency and hence the degree of inhibition of reverse transcription. © 1997 Elsevier Science B.V. All rights reserved.</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords class="keyword"><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text>HIV-1/HIV-2</ce:text></ce:keyword><ce:keyword><ce:text>Replication</ce:text></ce:keyword><ce:keyword><ce:text>Reverse transcription</ce:text></ce:keyword><ce:keyword><ce:text>Inhibition</ce:text></ce:keyword><ce:keyword><ce:text>Antisense</ce:text></ce:keyword><ce:keyword><ce:text>Trans-activating region</ce:text></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>Specific inhibition of in vitro reverse transcription using antisense oligonucleotides targeted to the TAR regions of HIV-1 and HIV-2</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>Specific inhibition of in vitro reverse transcription using antisense oligonucleotides targeted to the TAR regions of HIV-1 and HIV-2</title></titleInfo><name type="personal"><namePart type="given">Florence</namePart><namePart type="family">Boulme</namePart><affiliation>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Maritta</namePart><namePart type="family">Perälä-Heape</namePart><affiliation>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</affiliation><description>Present address: Fibrogen Europe, Kiviharjunke 11, 90220 Oulu, Finland.</description><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Leila</namePart><namePart type="family">Sarih-Cottin</namePart><affiliation>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Simon</namePart><namePart type="family">Litvak</namePart><affiliation>Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, 1, rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France</affiliation><description>Corresponding author. Fax: +33 556999057; E-mail: simon.litvak@ibgc.u-bordeaux2.fr</description><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1997</dateIssued><dateModified encoding="w3cdtf">1997-01-28</dateModified><copyrightDate encoding="w3cdtf">1997</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Abstract: Antisense oligonucleotides (ODNs) overlapping the stem-loop structure of the trans-activating responsive (TAR) element at the 5′ end of HIV-1 and HIV-2 viral RNAs were tested for their inhibitory effect on cDNA synthesis by HIV-1 and HIV-2 reverse transcriptases (RT). Inhibition of reverse transcription is sequence-specific and enhanced by the presence of the RT-associated RNase H activity. The degree of inhibition obtained with the anti-TAR antisense is significantly higher than with other HIV-1 targeted antisense ODNs used before [1]. Gel retardation showed a stable specific complex between the 16- and 25-mer anti-TAR HIV-1 selected ODNs and the target region. No complex was observed with a non-inhibitor 22-mer anti-TAR ODN and with the corresponding control sequences. Targeting of the first stem-loop in the 5′ region of HIV-2 RNA by anti-TAR ODNs inhibited very strongly reverse transcription by HIV-2 RT. The structure of the antisense and the target sequence affect annealing efficiency and hence the degree of inhibition of reverse transcription. © 1997 Elsevier Science B.V. All rights reserved.</abstract><note type="content">Section title: Rapid report</note><note type="content">Fig. 1: A: Antisense oligonucleotides (25-, 22- and 16-mers) directed against the HIV-1 TAR element are shown covering different regions of the stem-loop of the TAR RNA secondary structure. B: Schematic representation of the expected sizes of the HIV-1 cDNA products synthesized by HIV-1 RT from an anti-PBS primer in the presence of anti-TAR oligonucleotides of different lengths.</note><note type="content">Fig. 2: Effect of HIV-1 anti-TAR antisense ODNs on the initiation of reverse transcription. cDNA synthesis was performed either by HIV-RT (RNase H−), or (RNase H+) expressed and purified as described before [16–18]. M is the lane corresponding to the size markers in nucleotides. A: cDNA synthesis performed by HIV-1 RT (RNase H−), in the presence of anti-PBS 18-mer primer. Lane 1, control (cDNA synthesis in the presence of anti-PBS); lanes 2 and 5, anti-TAR 25-mer, respectively non-blocked (n) or blocked (b) in its 3′ end; lanes 3 and 6, anti-TAR 22-mer n and b; lanes 4 and 7, anti-TAR 16-mer n and b; antisense ODNs are used at a concentration of 1 μM. B: cDNA synthesis by HIV-RT p66/p51 (RNase H+) in the presence of anti-PBS 18-mer primer. Lane 1, control; lanes 2, 3 and 4, anti-TAR 25-mer at 0.5, 1.0 and 5.0 μM; lane 5, anti-TAR 22-mer at 1 μM; lanes 6, 7 and 8, anti-TAR 16-mer at 0.5, 1.0 and 5.0 μM, respectively. C: Effect of different concentrations of anti-TAR 25-mer antisense ODNs on the initiation of reverse transcription performed by HIV-RT (RNase H−) (□), or p66/p51 (RNase H+) (■).</note><note type="content">Fig. 3: Gel mobility shift of antisense/RNA target duplexes. Conditions are described in the text. Lane C: control labelled RNA alone; primer: anti-PBS (18-mer); lanes 1 and 2: anti-TAR (16-mer); lanes 3 and 4: anti-TAR (22-mer); lanes 5 and 6: anti-TAR (25-mer); lanes 7 and 8: mismatched anti-TAR (25-mer); lanes 9 and 10: scrambled anti-TAR (25-mer). Lanes 1, 3, 5, 7 and 9: 0,5 μM; lanes 2, 4, 6, 8 and 10: 1 μM. Experimental details are described in the text.</note><subject><genre>Keywords</genre><topic>HIV-1/HIV-2</topic><topic>Replication</topic><topic>Reverse transcription</topic><topic>Inhibition</topic><topic>Antisense</topic><topic>Trans-activating region</topic></subject><relatedItem type="host"><titleInfo><title>BBA - Gene Structure and Expression</title></titleInfo><titleInfo type="abbreviated"><title>BBAEXP</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1997</dateIssued></originInfo><identifier type="ISSN">0167-4781</identifier><identifier type="PII">S0167-4781(00)X0033-4</identifier><part><date>1997</date><detail type="volume"><number>1351</number><caption>vol.</caption></detail><detail type="issue"><number>3</number><caption>no.</caption></detail><extent unit="issue-pages"><start>249</start><end>364</end></extent><extent unit="pages"><start>249</start><end>255</end></extent></part></relatedItem><identifier type="istex">191164A3CCA1C0429CDAB785CEE915EA596FE97E</identifier><identifier type="ark">ark:/67375/6H6-06V08SB7-Z</identifier><identifier type="DOI">10.1016/S0167-4781(97)00026-2</identifier><identifier type="PII">S0167-4781(97)00026-2</identifier><accessCondition type="use and reproduction" contentType="copyright">©1997 Elsevier Science B.V.</accessCondition><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</recordContentSource><recordOrigin>Elsevier Science B.V., ©1997</recordOrigin></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-06V08SB7-Z/record.json</uri></json:item></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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