Use of interfering RNAs targeted against feline herpesvirus 1 glycoprotein D for inhibition of feline herpesvirus 1 infection of feline kidney cells
To evaluate the use of RNA interference targeted against feline herpesvirus 1 (FHV-1) glycoprotein D for inhibition of FHV-1 infection of feline kidney cells. Crandell-Rees feline kidney cells. Crandell-Rees feline kidney cells were transfected with small interfering RNAs (siRNAs) that were designed...
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| Veröffentlicht in: | American journal of veterinary research 2009-08, Vol.70 (8), p.1018-1025 |
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| creator | Wilkes, Rebbeca P Kania, Stephen A |
| description | To evaluate the use of RNA interference targeted against feline herpesvirus 1 (FHV-1) glycoprotein D for inhibition of FHV-1 infection of feline kidney cells.
Crandell-Rees feline kidney cells.
Crandell-Rees feline kidney cells were transfected with small interfering RNAs (siRNAs) that were designed to inhibit expression of FHV-1 glycoprotein D. The effectiveness of the treatment was determined via measurement of amounts of glycoprotein D mRNA, intracellular glycoprotein D, and glycoprotein D expressed on the surface of infected cells and comparison with appropriate control sample data.
2 of 6 siRNAs tested were highly effective in reducing expression (ie, knockdown) of glycoprotein D mRNA; there were 77% and 85% reductions in mRNA in treated samples, compared with findings in the control samples. The knockdown of glycoprotein D mRNA resulted in reduced glycoprotein D protein production, as evidenced by 27% and 43% decreases in expression of glycoprotein D on the surface of siRNA-treated, FHV-1-infected cells and decreased expression of the protein within infected cells, compared with control samples. Treatment with these siRNAs also resulted in inhibition of FHV-1 replication, with reductions of 84% and 77% in amounts of virus released into cell culture supernatant, compared with findings in control samples.
2 chemically produced siRNAs that targeted the glycoprotein D gene significantly reduced FHV-1 titers in treated cells, suggesting that glycoprotein D is necessary for production of infective virions. This gene is a potential target for RNA interference as a means of inhibition of FHV-1 infection of feline cells. |
| doi_str_mv | 10.2460/ajvr.70.8.1018 |
| format | Article |
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Crandell-Rees feline kidney cells.
Crandell-Rees feline kidney cells were transfected with small interfering RNAs (siRNAs) that were designed to inhibit expression of FHV-1 glycoprotein D. The effectiveness of the treatment was determined via measurement of amounts of glycoprotein D mRNA, intracellular glycoprotein D, and glycoprotein D expressed on the surface of infected cells and comparison with appropriate control sample data.
2 of 6 siRNAs tested were highly effective in reducing expression (ie, knockdown) of glycoprotein D mRNA; there were 77% and 85% reductions in mRNA in treated samples, compared with findings in the control samples. The knockdown of glycoprotein D mRNA resulted in reduced glycoprotein D protein production, as evidenced by 27% and 43% decreases in expression of glycoprotein D on the surface of siRNA-treated, FHV-1-infected cells and decreased expression of the protein within infected cells, compared with control samples. Treatment with these siRNAs also resulted in inhibition of FHV-1 replication, with reductions of 84% and 77% in amounts of virus released into cell culture supernatant, compared with findings in control samples.
2 chemically produced siRNAs that targeted the glycoprotein D gene significantly reduced FHV-1 titers in treated cells, suggesting that glycoprotein D is necessary for production of infective virions. This gene is a potential target for RNA interference as a means of inhibition of FHV-1 infection of feline cells.</description><identifier>ISSN: 0002-9645</identifier><identifier>EISSN: 1943-5681</identifier><identifier>DOI: 10.2460/ajvr.70.8.1018</identifier><identifier>PMID: 19645584</identifier><language>eng</language><publisher>United States</publisher><subject>Analysis of Variance ; Animals ; Blotting, Western - veterinary ; Cats ; Cell Line ; cell lines ; cultured cells ; disease control ; DNA Primers - genetics ; Felid herpesvirus 1 ; Feline herpesvirus 1 ; Flow Cytometry - veterinary ; gene expression ; Gene Expression Regulation - genetics ; glycoproteins ; Hemagglutination Tests - veterinary ; in vitro studies ; kidney cells ; kidneys ; messenger RNA ; protein synthesis ; Reverse Transcriptase Polymerase Chain Reaction - veterinary ; RNA Interference ; RNA, Small Interfering - genetics ; RNA, Small Interfering - pharmacology ; small interfering RNA ; Varicellovirus - genetics ; Viral Envelope Proteins - antagonists & inhibitors ; Viral Envelope Proteins - genetics ; Viral Plaque Assay - veterinary ; viral proteins ; Virus Attachment - drug effects ; virus replication</subject><ispartof>American journal of veterinary research, 2009-08, Vol.70 (8), p.1018-1025</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c389t-4c703dbd9357e4373cda88cea8b828011b70b64413ed76835e33dc313e47bd133</citedby><cites>FETCH-LOGICAL-c389t-4c703dbd9357e4373cda88cea8b828011b70b64413ed76835e33dc313e47bd133</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19645584$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wilkes, Rebbeca P</creatorcontrib><creatorcontrib>Kania, Stephen A</creatorcontrib><title>Use of interfering RNAs targeted against feline herpesvirus 1 glycoprotein D for inhibition of feline herpesvirus 1 infection of feline kidney cells</title><title>American journal of veterinary research</title><addtitle>Am J Vet Res</addtitle><description>To evaluate the use of RNA interference targeted against feline herpesvirus 1 (FHV-1) glycoprotein D for inhibition of FHV-1 infection of feline kidney cells.
Crandell-Rees feline kidney cells.
Crandell-Rees feline kidney cells were transfected with small interfering RNAs (siRNAs) that were designed to inhibit expression of FHV-1 glycoprotein D. The effectiveness of the treatment was determined via measurement of amounts of glycoprotein D mRNA, intracellular glycoprotein D, and glycoprotein D expressed on the surface of infected cells and comparison with appropriate control sample data.
2 of 6 siRNAs tested were highly effective in reducing expression (ie, knockdown) of glycoprotein D mRNA; there were 77% and 85% reductions in mRNA in treated samples, compared with findings in the control samples. The knockdown of glycoprotein D mRNA resulted in reduced glycoprotein D protein production, as evidenced by 27% and 43% decreases in expression of glycoprotein D on the surface of siRNA-treated, FHV-1-infected cells and decreased expression of the protein within infected cells, compared with control samples. Treatment with these siRNAs also resulted in inhibition of FHV-1 replication, with reductions of 84% and 77% in amounts of virus released into cell culture supernatant, compared with findings in control samples.
2 chemically produced siRNAs that targeted the glycoprotein D gene significantly reduced FHV-1 titers in treated cells, suggesting that glycoprotein D is necessary for production of infective virions. This gene is a potential target for RNA interference as a means of inhibition of FHV-1 infection of feline cells.</description><subject>Analysis of Variance</subject><subject>Animals</subject><subject>Blotting, Western - veterinary</subject><subject>Cats</subject><subject>Cell Line</subject><subject>cell lines</subject><subject>cultured cells</subject><subject>disease control</subject><subject>DNA Primers - genetics</subject><subject>Felid herpesvirus 1</subject><subject>Feline herpesvirus 1</subject><subject>Flow Cytometry - veterinary</subject><subject>gene expression</subject><subject>Gene Expression Regulation - genetics</subject><subject>glycoproteins</subject><subject>Hemagglutination Tests - veterinary</subject><subject>in vitro studies</subject><subject>kidney cells</subject><subject>kidneys</subject><subject>messenger RNA</subject><subject>protein synthesis</subject><subject>Reverse Transcriptase Polymerase Chain Reaction - veterinary</subject><subject>RNA Interference</subject><subject>RNA, Small Interfering - genetics</subject><subject>RNA, Small Interfering - pharmacology</subject><subject>small interfering RNA</subject><subject>Varicellovirus - genetics</subject><subject>Viral Envelope Proteins - antagonists & inhibitors</subject><subject>Viral Envelope Proteins - genetics</subject><subject>Viral Plaque Assay - veterinary</subject><subject>viral proteins</subject><subject>Virus Attachment - drug effects</subject><subject>virus replication</subject><issn>0002-9645</issn><issn>1943-5681</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU1r3DAQhkVpabZprz22OrUnuyPrc48h_YSQQNs9C1kaO0q98lbyBvZ_5AfHZhcKpfQ0zPDMwwwvIa8Z1I1Q8MHd3edaQ21qBsw8ISu2FrySyrCnZAUATbVWQp6RF6XcAbDGMPmcnLFlKI1YkYdNQTp2NKYJc4c5pp5-v74odHK5xwkDdb2LqUy0wyEmpLeYd1juY94Xymg_HPy4y-OEMdGPtBvzbLqNbZzimBbvP7di6tD_RfyKIeGBehyG8pI869xQ8NWpnpPN508_L79WVzdfvl1eXFWem_VUCa-BhzasudQouOY-OGM8OtOaxgBjrYZWCcE4Bq0Ml8h58HxuhW4D4_ycvD965w9-77FMdhvLcoFLOO6L1VIyAWCamXz3X1JpKaSRMIP1EfR5LCVjZ3c5bl0-WAZ2ScwuiVkN1tglsXnhzcm8b7cY_uCniGbg7RHo3Ghdn2Oxmx8NMA5MKQ4K-CM7XJ1F</recordid><startdate>20090801</startdate><enddate>20090801</enddate><creator>Wilkes, Rebbeca P</creator><creator>Kania, Stephen A</creator><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7TM</scope><scope>7U9</scope><scope>H94</scope></search><sort><creationdate>20090801</creationdate><title>Use of interfering RNAs targeted against feline herpesvirus 1 glycoprotein D for inhibition of feline herpesvirus 1 infection of feline kidney cells</title><author>Wilkes, Rebbeca P ; Kania, Stephen A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c389t-4c703dbd9357e4373cda88cea8b828011b70b64413ed76835e33dc313e47bd133</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Analysis of Variance</topic><topic>Animals</topic><topic>Blotting, Western - veterinary</topic><topic>Cats</topic><topic>Cell Line</topic><topic>cell lines</topic><topic>cultured cells</topic><topic>disease control</topic><topic>DNA Primers - genetics</topic><topic>Felid herpesvirus 1</topic><topic>Feline herpesvirus 1</topic><topic>Flow Cytometry - veterinary</topic><topic>gene expression</topic><topic>Gene Expression Regulation - genetics</topic><topic>glycoproteins</topic><topic>Hemagglutination Tests - veterinary</topic><topic>in vitro studies</topic><topic>kidney cells</topic><topic>kidneys</topic><topic>messenger RNA</topic><topic>protein synthesis</topic><topic>Reverse Transcriptase Polymerase Chain Reaction - veterinary</topic><topic>RNA Interference</topic><topic>RNA, Small Interfering - genetics</topic><topic>RNA, Small Interfering - pharmacology</topic><topic>small interfering RNA</topic><topic>Varicellovirus - genetics</topic><topic>Viral Envelope Proteins - antagonists & inhibitors</topic><topic>Viral Envelope Proteins - genetics</topic><topic>Viral Plaque Assay - veterinary</topic><topic>viral proteins</topic><topic>Virus Attachment - drug effects</topic><topic>virus replication</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wilkes, Rebbeca P</creatorcontrib><creatorcontrib>Kania, Stephen A</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><jtitle>American journal of veterinary research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wilkes, Rebbeca P</au><au>Kania, Stephen A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Use of interfering RNAs targeted against feline herpesvirus 1 glycoprotein D for inhibition of feline herpesvirus 1 infection of feline kidney cells</atitle><jtitle>American journal of veterinary research</jtitle><addtitle>Am J Vet Res</addtitle><date>2009-08-01</date><risdate>2009</risdate><volume>70</volume><issue>8</issue><spage>1018</spage><epage>1025</epage><pages>1018-1025</pages><issn>0002-9645</issn><eissn>1943-5681</eissn><abstract>To evaluate the use of RNA interference targeted against feline herpesvirus 1 (FHV-1) glycoprotein D for inhibition of FHV-1 infection of feline kidney cells.
Crandell-Rees feline kidney cells.
Crandell-Rees feline kidney cells were transfected with small interfering RNAs (siRNAs) that were designed to inhibit expression of FHV-1 glycoprotein D. The effectiveness of the treatment was determined via measurement of amounts of glycoprotein D mRNA, intracellular glycoprotein D, and glycoprotein D expressed on the surface of infected cells and comparison with appropriate control sample data.
2 of 6 siRNAs tested were highly effective in reducing expression (ie, knockdown) of glycoprotein D mRNA; there were 77% and 85% reductions in mRNA in treated samples, compared with findings in the control samples. The knockdown of glycoprotein D mRNA resulted in reduced glycoprotein D protein production, as evidenced by 27% and 43% decreases in expression of glycoprotein D on the surface of siRNA-treated, FHV-1-infected cells and decreased expression of the protein within infected cells, compared with control samples. Treatment with these siRNAs also resulted in inhibition of FHV-1 replication, with reductions of 84% and 77% in amounts of virus released into cell culture supernatant, compared with findings in control samples.
2 chemically produced siRNAs that targeted the glycoprotein D gene significantly reduced FHV-1 titers in treated cells, suggesting that glycoprotein D is necessary for production of infective virions. This gene is a potential target for RNA interference as a means of inhibition of FHV-1 infection of feline cells.</abstract><cop>United States</cop><pmid>19645584</pmid><doi>10.2460/ajvr.70.8.1018</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Analysis of Variance Animals Blotting, Western - veterinary Cats Cell Line cell lines cultured cells disease control DNA Primers - genetics Felid herpesvirus 1 Feline herpesvirus 1 Flow Cytometry - veterinary gene expression Gene Expression Regulation - genetics glycoproteins Hemagglutination Tests - veterinary in vitro studies kidney cells kidneys messenger RNA protein synthesis Reverse Transcriptase Polymerase Chain Reaction - veterinary RNA Interference RNA, Small Interfering - genetics RNA, Small Interfering - pharmacology small interfering RNA Varicellovirus - genetics Viral Envelope Proteins - antagonists & inhibitors Viral Envelope Proteins - genetics Viral Plaque Assay - veterinary viral proteins Virus Attachment - drug effects virus replication |
| title | Use of interfering RNAs targeted against feline herpesvirus 1 glycoprotein D for inhibition of feline herpesvirus 1 infection of feline kidney cells |
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