A novel RNA silencing vector to improve antigen expression and stability in Chinese hamster ovary cells
Abstract Chinese hamster ovary (CHO) cells and dihydrofolate reductase ( dhfr )/methotrexate (MTX) gene amplification system are routinely used to generate stable producer CHO cell clones in biopharmaceutical industries. The present study proposes a novel method by the co-amplification of the silenc...
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| Veröffentlicht in: | Vaccine 2007-05, Vol.25 (20), p.4103-4111 |
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| creator | Hong, Willy W.L Wu, Suh-Chin |
| description | Abstract Chinese hamster ovary (CHO) cells and dihydrofolate reductase ( dhfr )/methotrexate (MTX) gene amplification system are routinely used to generate stable producer CHO cell clones in biopharmaceutical industries. The present study proposes a novel method by the co-amplification of the silencing vector targeted to dhfr gene for improvements of selecting high-producing clones in dhfr -deficient and wild-type CHO cells. Using the silencing vector also resulted in improving the stability of the recombinant protein expression in the absence of MTX in the CHO/dhFr− and wild-type CHO cells. This new method is proposed to generate highly expressed stable cell clones of both dhfr -deficient and wild-type CHO cells for recombinant antigen production. Utilization of the silencing vector designed in this study can improve antigen expression through dhfr -directed gene amplification in other dhfr -competent cell lines for vaccine development. |
| doi_str_mv | 10.1016/j.vaccine.2007.02.012 |
| format | Article |
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The present study proposes a novel method by the co-amplification of the silencing vector targeted to dhfr gene for improvements of selecting high-producing clones in dhfr -deficient and wild-type CHO cells. Using the silencing vector also resulted in improving the stability of the recombinant protein expression in the absence of MTX in the CHO/dhFr− and wild-type CHO cells. This new method is proposed to generate highly expressed stable cell clones of both dhfr -deficient and wild-type CHO cells for recombinant antigen production. Utilization of the silencing vector designed in this study can improve antigen expression through dhfr -directed gene amplification in other dhfr -competent cell lines for vaccine development.</description><identifier>ISSN: 0264-410X</identifier><identifier>EISSN: 1873-2518</identifier><identifier>DOI: 10.1016/j.vaccine.2007.02.012</identifier><identifier>PMID: 17428585</identifier><identifier>CODEN: VACCDE</identifier><language>eng</language><publisher>Oxford: Elsevier Ltd</publisher><subject>Allergy and Immunology ; Animals ; Antigen ; Antigens ; Antigens - biosynthesis ; Antigens - genetics ; Applied microbiology ; Base Sequence ; Biological and medical sciences ; CHO Cells ; Clone Cells ; Cloning ; Cricetinae ; Cricetulus ; Cytomegalovirus ; Deoxyribonucleic acid ; Dihydrofolate reductase ; DNA ; Fundamental and applied biological sciences. Psychology ; Gene Amplification ; Gene expression ; Genetic Vectors - genetics ; Green Fluorescent Proteins ; Methotrexate - pharmacology ; Mice ; Microbiology ; Molecular Sequence Data ; Plasmids ; Proteins ; RNA Interference ; RNA silencing ; Rodents ; Tetrahydrofolate Dehydrogenase - biosynthesis ; Tetrahydrofolate Dehydrogenase - deficiency ; Tetrahydrofolate Dehydrogenase - genetics ; Transfection ; Vaccines ; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects)</subject><ispartof>Vaccine, 2007-05, Vol.25 (20), p.4103-4111</ispartof><rights>Elsevier Ltd</rights><rights>2007 Elsevier Ltd</rights><rights>2007 INIST-CNRS</rights><rights>Copyright Elsevier Limited May 16, 2007</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c507t-21ac2f7c84750a8cb3749ab8f7a854528daeb679dfd0d82bcfffdb9eaf1120343</citedby><cites>FETCH-LOGICAL-c507t-21ac2f7c84750a8cb3749ab8f7a854528daeb679dfd0d82bcfffdb9eaf1120343</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.proquest.com/docview/1559084171?pq-origsite=primo$$EHTML$$P50$$Gproquest$$H</linktohtml><link.rule.ids>315,782,786,3552,27931,27932,46002,64392,64394,64396,72476</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18739291$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17428585$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hong, Willy W.L</creatorcontrib><creatorcontrib>Wu, Suh-Chin</creatorcontrib><title>A novel RNA silencing vector to improve antigen expression and stability in Chinese hamster ovary cells</title><title>Vaccine</title><addtitle>Vaccine</addtitle><description>Abstract Chinese hamster ovary (CHO) cells and dihydrofolate reductase ( dhfr )/methotrexate (MTX) gene amplification system are routinely used to generate stable producer CHO cell clones in biopharmaceutical industries. The present study proposes a novel method by the co-amplification of the silencing vector targeted to dhfr gene for improvements of selecting high-producing clones in dhfr -deficient and wild-type CHO cells. Using the silencing vector also resulted in improving the stability of the recombinant protein expression in the absence of MTX in the CHO/dhFr− and wild-type CHO cells. This new method is proposed to generate highly expressed stable cell clones of both dhfr -deficient and wild-type CHO cells for recombinant antigen production. Utilization of the silencing vector designed in this study can improve antigen expression through dhfr -directed gene amplification in other dhfr -competent cell lines for vaccine development.</description><subject>Allergy and Immunology</subject><subject>Animals</subject><subject>Antigen</subject><subject>Antigens</subject><subject>Antigens - biosynthesis</subject><subject>Antigens - genetics</subject><subject>Applied microbiology</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>CHO Cells</subject><subject>Clone Cells</subject><subject>Cloning</subject><subject>Cricetinae</subject><subject>Cricetulus</subject><subject>Cytomegalovirus</subject><subject>Deoxyribonucleic acid</subject><subject>Dihydrofolate reductase</subject><subject>DNA</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Amplification</subject><subject>Gene expression</subject><subject>Genetic Vectors - genetics</subject><subject>Green Fluorescent Proteins</subject><subject>Methotrexate - pharmacology</subject><subject>Mice</subject><subject>Microbiology</subject><subject>Molecular Sequence Data</subject><subject>Plasmids</subject><subject>Proteins</subject><subject>RNA Interference</subject><subject>RNA silencing</subject><subject>Rodents</subject><subject>Tetrahydrofolate Dehydrogenase - biosynthesis</subject><subject>Tetrahydrofolate Dehydrogenase - deficiency</subject><subject>Tetrahydrofolate Dehydrogenase - genetics</subject><subject>Transfection</subject><subject>Vaccines</subject><subject>Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects)</subject><issn>0264-410X</issn><issn>1873-2518</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqFkl2L1DAUhoMo7rj6E5SA6F3rSZo06Y0yDH7BouAHeBfS9HQ2Y6edTTrF-femTGFgb_YqUJ7z5j19QshLBjkDVr7b5ZN1zveYcwCVA8-B8UdkxbQqMi6ZfkxWwEuRCQZ_rsizGHcAIAtWPSVXTAmupZYrsl3Tfpiwoz--rWn0HfYpc0sndOMQ6DhQvz-EBFDbj36LPcV_h4Ax-qFPnxoaR1v7zo8n6nu6uU19ItJbu48jBjpMNpyow66Lz8mT1nYRXyznNfn96eOvzZfs5vvnr5v1TeYkqDHjzDreKqeFkmC1qwslKlvrVlktheS6sViXqmraBhrNa9e2bVNXaFvGOBSiuCZvz7mp9d0R42j2Ps4NbI_DMRoFouBlJR8EUxoXEiCBr--Bu-EY-rSEYVJWoAVTLFHyTLkwxBiwNYfg92l9w8DMwszOLMLMLMwAN0lYmnu1pB_rPTaXqcVQAt4sgI3Odm2wyVC8cEl3xau5wIczh-nvTh6Dic4nm9j4kGSaZvAPVnl_L8F1vvfp0r94wnjZ2sQ0YH7Or2t-XKAAmCpF8R8d1svf</recordid><startdate>20070516</startdate><enddate>20070516</enddate><creator>Hong, Willy W.L</creator><creator>Wu, Suh-Chin</creator><general>Elsevier Ltd</general><general>Elsevier</general><general>Elsevier Limited</general><scope>IQODW</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>3V.</scope><scope>7QL</scope><scope>7RV</scope><scope>7T2</scope><scope>7T5</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88C</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9-</scope><scope>K9.</scope><scope>KB0</scope><scope>LK8</scope><scope>M0R</scope><scope>M0S</scope><scope>M0T</scope><scope>M1P</scope><scope>M2O</scope><scope>M7N</scope><scope>M7P</scope><scope>MBDVC</scope><scope>NAPCQ</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7QO</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20070516</creationdate><title>A novel RNA silencing vector to improve antigen expression and stability in Chinese hamster ovary cells</title><author>Hong, Willy W.L ; Wu, Suh-Chin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c507t-21ac2f7c84750a8cb3749ab8f7a854528daeb679dfd0d82bcfffdb9eaf1120343</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Allergy and Immunology</topic><topic>Animals</topic><topic>Antigen</topic><topic>Antigens</topic><topic>Antigens - biosynthesis</topic><topic>Antigens - genetics</topic><topic>Applied microbiology</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>CHO Cells</topic><topic>Clone Cells</topic><topic>Cloning</topic><topic>Cricetinae</topic><topic>Cricetulus</topic><topic>Cytomegalovirus</topic><topic>Deoxyribonucleic acid</topic><topic>Dihydrofolate reductase</topic><topic>DNA</topic><topic>Fundamental and applied biological sciences. 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Academic</collection><jtitle>Vaccine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hong, Willy W.L</au><au>Wu, Suh-Chin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A novel RNA silencing vector to improve antigen expression and stability in Chinese hamster ovary cells</atitle><jtitle>Vaccine</jtitle><addtitle>Vaccine</addtitle><date>2007-05-16</date><risdate>2007</risdate><volume>25</volume><issue>20</issue><spage>4103</spage><epage>4111</epage><pages>4103-4111</pages><issn>0264-410X</issn><eissn>1873-2518</eissn><coden>VACCDE</coden><abstract>Abstract Chinese hamster ovary (CHO) cells and dihydrofolate reductase ( dhfr )/methotrexate (MTX) gene amplification system are routinely used to generate stable producer CHO cell clones in biopharmaceutical industries. The present study proposes a novel method by the co-amplification of the silencing vector targeted to dhfr gene for improvements of selecting high-producing clones in dhfr -deficient and wild-type CHO cells. Using the silencing vector also resulted in improving the stability of the recombinant protein expression in the absence of MTX in the CHO/dhFr− and wild-type CHO cells. This new method is proposed to generate highly expressed stable cell clones of both dhfr -deficient and wild-type CHO cells for recombinant antigen production. Utilization of the silencing vector designed in this study can improve antigen expression through dhfr -directed gene amplification in other dhfr -competent cell lines for vaccine development.</abstract><cop>Oxford</cop><pub>Elsevier Ltd</pub><pmid>17428585</pmid><doi>10.1016/j.vaccine.2007.02.012</doi><tpages>9</tpages></addata></record> |
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| subjects | Allergy and Immunology Animals Antigen Antigens Antigens - biosynthesis Antigens - genetics Applied microbiology Base Sequence Biological and medical sciences CHO Cells Clone Cells Cloning Cricetinae Cricetulus Cytomegalovirus Deoxyribonucleic acid Dihydrofolate reductase DNA Fundamental and applied biological sciences. Psychology Gene Amplification Gene expression Genetic Vectors - genetics Green Fluorescent Proteins Methotrexate - pharmacology Mice Microbiology Molecular Sequence Data Plasmids Proteins RNA Interference RNA silencing Rodents Tetrahydrofolate Dehydrogenase - biosynthesis Tetrahydrofolate Dehydrogenase - deficiency Tetrahydrofolate Dehydrogenase - genetics Transfection Vaccines Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects) |
| title | A novel RNA silencing vector to improve antigen expression and stability in Chinese hamster ovary cells |
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