Efficient gene silencing by delivery of locked nucleic acid antisense oligonucleotides, unassisted by transfection reagents.
Identifieur interne : 001F87 ( PubMed/Corpus ); précédent : 001F86; suivant : 001F88Efficient gene silencing by delivery of locked nucleic acid antisense oligonucleotides, unassisted by transfection reagents.
Auteurs : C A Stein ; J Bo Hansen ; Johnathan Lai ; Sijian Wu ; Anatoliy Voskresenskiy ; Anja H G ; Jesper Worm ; Maj Hedtj Rn ; Naira Souleimanian ; Paul Miller ; Harris S. Soifer ; Daniella Castanotto ; Luba Benimetskaya ; Henrik Rum ; Troels KochSource :
- Nucleic acids research [ 1362-4962 ] ; 2010.
English descriptors
- KwdEn :
- Animals, Cell Line, Tumor, Gene Silencing, Humans, Indicators and Reagents, Mice, Oligonucleotides (administration & dosage), Oligonucleotides (analysis), Oligonucleotides, Antisense (administration & dosage), Oligonucleotides, Antisense (analysis), Proto-Oncogene Proteins c-bcl-2 (genetics), Proto-Oncogene Proteins c-bcl-2 (metabolism), Transfection.
- MESH :
- chemical , administration & dosage : Oligonucleotides, Oligonucleotides, Antisense.
- chemical , analysis : Oligonucleotides, Oligonucleotides, Antisense.
- chemical , genetics : Proto-Oncogene Proteins c-bcl-2.
- chemical , metabolism : Proto-Oncogene Proteins c-bcl-2.
- chemical : Indicators and Reagents.
- Animals, Cell Line, Tumor, Gene Silencing, Humans, Mice, Transfection.
Abstract
For the past 15-20 years, the intracellular delivery and silencing activity of oligodeoxynucleotides have been essentially completely dependent on the use of a delivery technology (e.g. lipofection). We have developed a method (called 'gymnosis') that does not require the use of any transfection reagent or any additives to serum whatsoever, but rather takes advantage of the normal growth properties of cells in tissue culture in order to promote productive oligonucleotide uptake. This robust method permits the sequence-specific silencing of multiple targets in a large number of cell types in tissue culture, both at the protein and mRNA level, at concentrations in the low micromolar range. Optimum results were obtained with locked nucleic acid (LNA) phosphorothioate gap-mers. By appropriate manipulation of oligonucleotide dosing, this silencing can be continuously maintained with little or no toxicity for >240 days. High levels of oligonucleotide in the cell nucleus are not a requirement for gene silencing, contrary to long accepted dogma. In addition, gymnotic delivery can efficiently deliver oligonucleotides to suspension cells that are known to be very difficult to transfect. Finally, the pattern of gene silencing of in vitro gymnotically delivered oligonucleotides correlates particularly well with in vivo silencing. The establishment of this link is of particular significance to those in the academic research and drug discovery and development communities.
DOI: 10.1093/nar/gkp841
PubMed: 19854938
Links to Exploration step
pubmed:19854938Le document en format XML
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Soifer</name></author><author><name sortKey="Castanotto, Daniella" sort="Castanotto, Daniella" uniqKey="Castanotto D" first="Daniella" last="Castanotto">Daniella Castanotto</name></author><author><name sortKey="Benimetskaya, Luba" sort="Benimetskaya, Luba" uniqKey="Benimetskaya L" first="Luba" last="Benimetskaya">Luba Benimetskaya</name></author><author><name sortKey=" Rum, Henrik" sort=" Rum, Henrik" uniqKey=" Rum H" first="Henrik" last=" Rum">Henrik Rum</name></author><author><name sortKey="Koch, Troels" sort="Koch, Troels" uniqKey="Koch T" first="Troels" last="Koch">Troels Koch</name></author></analytic><series><title level="j">Nucleic acids research</title><idno type="eISSN">1362-4962</idno><imprint><date when="2010" type="published">2010</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Cell Line, Tumor</term><term>Gene Silencing</term><term>Humans</term><term>Indicators and Reagents</term><term>Mice</term><term>Oligonucleotides (administration & dosage)</term><term>Oligonucleotides (analysis)</term><term>Oligonucleotides, Antisense (administration & dosage)</term><term>Oligonucleotides, Antisense (analysis)</term><term>Proto-Oncogene Proteins c-bcl-2 (genetics)</term><term>Proto-Oncogene Proteins c-bcl-2 (metabolism)</term><term>Transfection</term></keywords><keywords scheme="MESH" type="chemical" qualifier="administration & dosage" xml:lang="en"><term>Oligonucleotides</term><term>Oligonucleotides, Antisense</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Oligonucleotides</term><term>Oligonucleotides, Antisense</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Proto-Oncogene Proteins c-bcl-2</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Proto-Oncogene Proteins c-bcl-2</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Indicators and Reagents</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cell Line, Tumor</term><term>Gene Silencing</term><term>Humans</term><term>Mice</term><term>Transfection</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">For the past 15-20 years, the intracellular delivery and silencing activity of oligodeoxynucleotides have been essentially completely dependent on the use of a delivery technology (e.g. lipofection). We have developed a method (called 'gymnosis') that does not require the use of any transfection reagent or any additives to serum whatsoever, but rather takes advantage of the normal growth properties of cells in tissue culture in order to promote productive oligonucleotide uptake. This robust method permits the sequence-specific silencing of multiple targets in a large number of cell types in tissue culture, both at the protein and mRNA level, at concentrations in the low micromolar range. Optimum results were obtained with locked nucleic acid (LNA) phosphorothioate gap-mers. By appropriate manipulation of oligonucleotide dosing, this silencing can be continuously maintained with little or no toxicity for >240 days. High levels of oligonucleotide in the cell nucleus are not a requirement for gene silencing, contrary to long accepted dogma. In addition, gymnotic delivery can efficiently deliver oligonucleotides to suspension cells that are known to be very difficult to transfect. Finally, the pattern of gene silencing of in vitro gymnotically delivered oligonucleotides correlates particularly well with in vivo silencing. The establishment of this link is of particular significance to those in the academic research and drug discovery and development communities.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">19854938</PMID><DateCompleted><Year>2010</Year><Month>02</Month><Day>12</Day></DateCompleted><DateRevised><Year>2018</Year><Month>11</Month><Day>13</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1362-4962</ISSN><JournalIssue CitedMedium="Internet"><Volume>38</Volume><Issue>1</Issue><PubDate><Year>2010</Year><Month>Jan</Month></PubDate></JournalIssue><Title>Nucleic acids research</Title><ISOAbbreviation>Nucleic Acids Res.</ISOAbbreviation></Journal><ArticleTitle>Efficient gene silencing by delivery of locked nucleic acid antisense oligonucleotides, unassisted by transfection reagents.</ArticleTitle><Pagination><MedlinePgn>e3</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1093/nar/gkp841</ELocationID><Abstract><AbstractText>For the past 15-20 years, the intracellular delivery and silencing activity of oligodeoxynucleotides have been essentially completely dependent on the use of a delivery technology (e.g. lipofection). We have developed a method (called 'gymnosis') that does not require the use of any transfection reagent or any additives to serum whatsoever, but rather takes advantage of the normal growth properties of cells in tissue culture in order to promote productive oligonucleotide uptake. This robust method permits the sequence-specific silencing of multiple targets in a large number of cell types in tissue culture, both at the protein and mRNA level, at concentrations in the low micromolar range. Optimum results were obtained with locked nucleic acid (LNA) phosphorothioate gap-mers. By appropriate manipulation of oligonucleotide dosing, this silencing can be continuously maintained with little or no toxicity for >240 days. High levels of oligonucleotide in the cell nucleus are not a requirement for gene silencing, contrary to long accepted dogma. In addition, gymnotic delivery can efficiently deliver oligonucleotides to suspension cells that are known to be very difficult to transfect. Finally, the pattern of gene silencing of in vitro gymnotically delivered oligonucleotides correlates particularly well with in vivo silencing. The establishment of this link is of particular significance to those in the academic research and drug discovery and development communities.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Stein</LastName><ForeName>C A</ForeName><Initials>CA</Initials><AffiliationInfo><Affiliation>Albert Einstein-Montefiore Cancer Center, Montefiore Medical Center, Bronx, NY 10467, USA. cstein@montefiore.org</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hansen</LastName><ForeName>J Bo</ForeName><Initials>JB</Initials></Author><Author ValidYN="Y"><LastName>Lai</LastName><ForeName>Johnathan</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Wu</LastName><ForeName>SiJian</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Voskresenskiy</LastName><ForeName>Anatoliy</ForeName><Initials>A</Initials></Author><Author ValidYN="Y"><LastName>Høg</LastName><ForeName>Anja</ForeName><Initials>A</Initials></Author><Author ValidYN="Y"><LastName>Worm</LastName><ForeName>Jesper</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Hedtjärn</LastName><ForeName>Maj</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Souleimanian</LastName><ForeName>Naira</ForeName><Initials>N</Initials></Author><Author ValidYN="Y"><LastName>Miller</LastName><ForeName>Paul</ForeName><Initials>P</Initials></Author><Author ValidYN="Y"><LastName>Soifer</LastName><ForeName>Harris S</ForeName><Initials>HS</Initials></Author><Author ValidYN="Y"><LastName>Castanotto</LastName><ForeName>Daniella</ForeName><Initials>D</Initials></Author><Author ValidYN="Y"><LastName>Benimetskaya</LastName><ForeName>Luba</ForeName><Initials>L</Initials></Author><Author ValidYN="Y"><LastName>Ørum</LastName><ForeName>Henrik</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Koch</LastName><ForeName>Troels</ForeName><Initials>T</Initials></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>R01 CA108415</GrantID><Acronym>CA</Acronym><Agency>NCI NIH HHS</Agency><Country>United States</Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D052061">Research Support, N.I.H., Extramural</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2009</Year><Month>10</Month><Day>23</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Nucleic Acids Res</MedlineTA><NlmUniqueID>0411011</NlmUniqueID><ISSNLinking>0305-1048</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D007202">Indicators and Reagents</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D009841">Oligonucleotides</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D016376">Oligonucleotides, Antisense</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D019253">Proto-Oncogene Proteins c-bcl-2</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C477371">locked nucleic acid</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D045744" MajorTopicYN="N">Cell Line, Tumor</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D020868" MajorTopicYN="Y">Gene Silencing</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007202" MajorTopicYN="N">Indicators and Reagents</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D051379" MajorTopicYN="N">Mice</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009841" MajorTopicYN="N">Oligonucleotides</DescriptorName><QualifierName UI="Q000008" MajorTopicYN="Y">administration & dosage</QualifierName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D016376" MajorTopicYN="N">Oligonucleotides, Antisense</DescriptorName><QualifierName UI="Q000008" MajorTopicYN="Y">administration & dosage</QualifierName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D019253" MajorTopicYN="N">Proto-Oncogene Proteins c-bcl-2</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014162" MajorTopicYN="N">Transfection</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2009</Year><Month>10</Month><Day>27</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2009</Year><Month>10</Month><Day>27</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2010</Year><Month>2</Month><Day>13</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">19854938</ArticleId><ArticleId IdType="pii">gkp841</ArticleId><ArticleId IdType="doi">10.1093/nar/gkp841</ArticleId><ArticleId IdType="pmc">PMC2800216</ArticleId></ArticleIdList><ReferenceList><Reference><Citation>J Drug Target. 2003 Jul;11(6):311-23</Citation><ArticleIdList><ArticleId 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