A genome-wide functional assay of signal transduction in living mammalian cells
We describe a genome-wide functional assay for rapid isolation of cell clones and genetic elements responsive to specific stimuli. A promoterless β-lactamase reporter gene was transfected into a human T-cell line to generate a living library of reporter-tagged clones. When loaded with a cell-permeab...
Gespeichert in:
| Veröffentlicht in: | Nature biotechnology 1998-12, Vol.16 (13), p.1329-1333 |
|---|---|
| Hauptverfasser: | , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 1333 |
|---|---|
| container_issue | 13 |
| container_start_page | 1329 |
| container_title | Nature biotechnology |
| container_volume | 16 |
| creator | Whitney, Mike Rockenstein, Edward Cantin, Greg Knapp, Tom Zlokarnik, Gregor Sanders, Pam Durick, Kyle Craig, Frank F. Negulescu, Paul A. |
| description | We describe a genome-wide functional assay for rapid isolation of cell clones and genetic elements responsive to specific stimuli. A promoterless β-lactamase reporter gene was transfected into a human T-cell line to generate a living library of reporter-tagged clones. When loaded with a cell-permeable fluorogenic substrate, the cell library simultaneously reports the expression of a large number of endogenous genes. Flow cytometry was used to recover individual clones whose reporter-tagged genes were either induced or repressed following T-cell activation. Responsive clones were expanded and analyzed pharmacologically to identify patterns of regulation associated with specific genes. Although demonstrated using T cells, the genomic assay could be applied to map downstream transcriptional consequences for any propagating cell line in response to any stimulus of interest. |
| doi_str_mv | 10.1038/4302 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_17119285</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>17119285</sourcerecordid><originalsourceid>FETCH-LOGICAL-c360t-da41b3c7cfb5896b8e524348f846f4b693f627ccb96bb32dbf1a6a76dff1fbc73</originalsourceid><addsrcrecordid>eNpVkF1LwzAUhoMoc3b-BCEX6l01adK0vRzDLxjsRq9LkiYlo01nTqvs39u64fAqJ7wvz0kehBaUPFDC8kfOSHKG5jTlIqaiEOfjTPIsJjQVl-gKYEsIEVyIGZoVecoEZXO0WeLa-K418berDLaD173rvGywBJB73FkMrp7ufZAequE3xs7jxn05X-NWtq1snPRYm6aBBbqwsgFzfTwj9PH89L56jdebl7fVch1rJkgfV5JTxXSmrUrzQqjcpAlnPLc5F5YrUTArkkxrNWaKJZWyVAqZicpaapXOWITuD9xd6D4HA33ZOpheIL3pBihpRmmRjL-M0O2hqEMHEIwtd8G1MuxLSsrJWzl5G2s3R96gWlP9lY6iTpidBC0bO8rQDk6sLOFFMmHuDjUYE1-bUG67IYz64P-6H0-dgK0</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>17119285</pqid></control><display><type>article</type><title>A genome-wide functional assay of signal transduction in living mammalian cells</title><source>MEDLINE</source><source>Springer Nature - Connect here FIRST to enable access</source><source>SpringerLink Journals - AutoHoldings</source><creator>Whitney, Mike ; Rockenstein, Edward ; Cantin, Greg ; Knapp, Tom ; Zlokarnik, Gregor ; Sanders, Pam ; Durick, Kyle ; Craig, Frank F. ; Negulescu, Paul A.</creator><creatorcontrib>Whitney, Mike ; Rockenstein, Edward ; Cantin, Greg ; Knapp, Tom ; Zlokarnik, Gregor ; Sanders, Pam ; Durick, Kyle ; Craig, Frank F. ; Negulescu, Paul A.</creatorcontrib><description>We describe a genome-wide functional assay for rapid isolation of cell clones and genetic elements responsive to specific stimuli. A promoterless β-lactamase reporter gene was transfected into a human T-cell line to generate a living library of reporter-tagged clones. When loaded with a cell-permeable fluorogenic substrate, the cell library simultaneously reports the expression of a large number of endogenous genes. Flow cytometry was used to recover individual clones whose reporter-tagged genes were either induced or repressed following T-cell activation. Responsive clones were expanded and analyzed pharmacologically to identify patterns of regulation associated with specific genes. Although demonstrated using T cells, the genomic assay could be applied to map downstream transcriptional consequences for any propagating cell line in response to any stimulus of interest.</description><identifier>ISSN: 1087-0156</identifier><identifier>EISSN: 1546-1696</identifier><identifier>DOI: 10.1038/4302</identifier><identifier>PMID: 9853613</identifier><identifier>CODEN: NABIF9</identifier><language>eng</language><publisher>New York: Nature Publishing Group US</publisher><subject>Agriculture ; beta-Lactamases - genetics ; Bioinformatics ; Biological and medical sciences ; Biomedical and Life Sciences ; Biomedical Engineering/Biotechnology ; Biomedicine ; Biotechnology ; Cell Separation ; Diverse techniques ; Enzyme Activation ; Flow Cytometry ; Fluorescent Dyes ; Fundamental and applied biological sciences. Psychology ; Genes, Reporter ; Genome ; Humans ; Jurkat Cells ; Life Sciences ; Molecular and cellular biology ; research-article ; Signal Transduction ; Tetradecanoylphorbol Acetate - pharmacology ; Thapsigargin - pharmacology</subject><ispartof>Nature biotechnology, 1998-12, Vol.16 (13), p.1329-1333</ispartof><rights>Nature America Inc. 1998</rights><rights>1999 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c360t-da41b3c7cfb5896b8e524348f846f4b693f627ccb96bb32dbf1a6a76dff1fbc73</citedby><cites>FETCH-LOGICAL-c360t-da41b3c7cfb5896b8e524348f846f4b693f627ccb96bb32dbf1a6a76dff1fbc73</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/4302$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/4302$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27903,27904,41467,42536,51298</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1724922$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9853613$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Whitney, Mike</creatorcontrib><creatorcontrib>Rockenstein, Edward</creatorcontrib><creatorcontrib>Cantin, Greg</creatorcontrib><creatorcontrib>Knapp, Tom</creatorcontrib><creatorcontrib>Zlokarnik, Gregor</creatorcontrib><creatorcontrib>Sanders, Pam</creatorcontrib><creatorcontrib>Durick, Kyle</creatorcontrib><creatorcontrib>Craig, Frank F.</creatorcontrib><creatorcontrib>Negulescu, Paul A.</creatorcontrib><title>A genome-wide functional assay of signal transduction in living mammalian cells</title><title>Nature biotechnology</title><addtitle>Nat Biotechnol</addtitle><addtitle>Nat Biotechnol</addtitle><description>We describe a genome-wide functional assay for rapid isolation of cell clones and genetic elements responsive to specific stimuli. A promoterless β-lactamase reporter gene was transfected into a human T-cell line to generate a living library of reporter-tagged clones. When loaded with a cell-permeable fluorogenic substrate, the cell library simultaneously reports the expression of a large number of endogenous genes. Flow cytometry was used to recover individual clones whose reporter-tagged genes were either induced or repressed following T-cell activation. Responsive clones were expanded and analyzed pharmacologically to identify patterns of regulation associated with specific genes. Although demonstrated using T cells, the genomic assay could be applied to map downstream transcriptional consequences for any propagating cell line in response to any stimulus of interest.</description><subject>Agriculture</subject><subject>beta-Lactamases - genetics</subject><subject>Bioinformatics</subject><subject>Biological and medical sciences</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedical Engineering/Biotechnology</subject><subject>Biomedicine</subject><subject>Biotechnology</subject><subject>Cell Separation</subject><subject>Diverse techniques</subject><subject>Enzyme Activation</subject><subject>Flow Cytometry</subject><subject>Fluorescent Dyes</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes, Reporter</subject><subject>Genome</subject><subject>Humans</subject><subject>Jurkat Cells</subject><subject>Life Sciences</subject><subject>Molecular and cellular biology</subject><subject>research-article</subject><subject>Signal Transduction</subject><subject>Tetradecanoylphorbol Acetate - pharmacology</subject><subject>Thapsigargin - pharmacology</subject><issn>1087-0156</issn><issn>1546-1696</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkF1LwzAUhoMoc3b-BCEX6l01adK0vRzDLxjsRq9LkiYlo01nTqvs39u64fAqJ7wvz0kehBaUPFDC8kfOSHKG5jTlIqaiEOfjTPIsJjQVl-gKYEsIEVyIGZoVecoEZXO0WeLa-K418berDLaD173rvGywBJB73FkMrp7ufZAequE3xs7jxn05X-NWtq1snPRYm6aBBbqwsgFzfTwj9PH89L56jdebl7fVch1rJkgfV5JTxXSmrUrzQqjcpAlnPLc5F5YrUTArkkxrNWaKJZWyVAqZicpaapXOWITuD9xd6D4HA33ZOpheIL3pBihpRmmRjL-M0O2hqEMHEIwtd8G1MuxLSsrJWzl5G2s3R96gWlP9lY6iTpidBC0bO8rQDk6sLOFFMmHuDjUYE1-bUG67IYz64P-6H0-dgK0</recordid><startdate>19981201</startdate><enddate>19981201</enddate><creator>Whitney, Mike</creator><creator>Rockenstein, Edward</creator><creator>Cantin, Greg</creator><creator>Knapp, Tom</creator><creator>Zlokarnik, Gregor</creator><creator>Sanders, Pam</creator><creator>Durick, Kyle</creator><creator>Craig, Frank F.</creator><creator>Negulescu, Paul A.</creator><general>Nature Publishing Group US</general><general>Nature</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>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>19981201</creationdate><title>A genome-wide functional assay of signal transduction in living mammalian cells</title><author>Whitney, Mike ; Rockenstein, Edward ; Cantin, Greg ; Knapp, Tom ; Zlokarnik, Gregor ; Sanders, Pam ; Durick, Kyle ; Craig, Frank F. ; Negulescu, Paul A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c360t-da41b3c7cfb5896b8e524348f846f4b693f627ccb96bb32dbf1a6a76dff1fbc73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Agriculture</topic><topic>beta-Lactamases - genetics</topic><topic>Bioinformatics</topic><topic>Biological and medical sciences</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedical Engineering/Biotechnology</topic><topic>Biomedicine</topic><topic>Biotechnology</topic><topic>Cell Separation</topic><topic>Diverse techniques</topic><topic>Enzyme Activation</topic><topic>Flow Cytometry</topic><topic>Fluorescent Dyes</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes, Reporter</topic><topic>Genome</topic><topic>Humans</topic><topic>Jurkat Cells</topic><topic>Life Sciences</topic><topic>Molecular and cellular biology</topic><topic>research-article</topic><topic>Signal Transduction</topic><topic>Tetradecanoylphorbol Acetate - pharmacology</topic><topic>Thapsigargin - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Whitney, Mike</creatorcontrib><creatorcontrib>Rockenstein, Edward</creatorcontrib><creatorcontrib>Cantin, Greg</creatorcontrib><creatorcontrib>Knapp, Tom</creatorcontrib><creatorcontrib>Zlokarnik, Gregor</creatorcontrib><creatorcontrib>Sanders, Pam</creatorcontrib><creatorcontrib>Durick, Kyle</creatorcontrib><creatorcontrib>Craig, Frank F.</creatorcontrib><creatorcontrib>Negulescu, Paul A.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Nature biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Whitney, Mike</au><au>Rockenstein, Edward</au><au>Cantin, Greg</au><au>Knapp, Tom</au><au>Zlokarnik, Gregor</au><au>Sanders, Pam</au><au>Durick, Kyle</au><au>Craig, Frank F.</au><au>Negulescu, Paul A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A genome-wide functional assay of signal transduction in living mammalian cells</atitle><jtitle>Nature biotechnology</jtitle><stitle>Nat Biotechnol</stitle><addtitle>Nat Biotechnol</addtitle><date>1998-12-01</date><risdate>1998</risdate><volume>16</volume><issue>13</issue><spage>1329</spage><epage>1333</epage><pages>1329-1333</pages><issn>1087-0156</issn><eissn>1546-1696</eissn><coden>NABIF9</coden><abstract>We describe a genome-wide functional assay for rapid isolation of cell clones and genetic elements responsive to specific stimuli. A promoterless β-lactamase reporter gene was transfected into a human T-cell line to generate a living library of reporter-tagged clones. When loaded with a cell-permeable fluorogenic substrate, the cell library simultaneously reports the expression of a large number of endogenous genes. Flow cytometry was used to recover individual clones whose reporter-tagged genes were either induced or repressed following T-cell activation. Responsive clones were expanded and analyzed pharmacologically to identify patterns of regulation associated with specific genes. Although demonstrated using T cells, the genomic assay could be applied to map downstream transcriptional consequences for any propagating cell line in response to any stimulus of interest.</abstract><cop>New York</cop><pub>Nature Publishing Group US</pub><pmid>9853613</pmid><doi>10.1038/4302</doi><tpages>5</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1087-0156 |
| ispartof | Nature biotechnology, 1998-12, Vol.16 (13), p.1329-1333 |
| issn | 1087-0156 1546-1696 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_17119285 |
| source | MEDLINE; Springer Nature - Connect here FIRST to enable access; SpringerLink Journals - AutoHoldings |
| subjects | Agriculture beta-Lactamases - genetics Bioinformatics Biological and medical sciences Biomedical and Life Sciences Biomedical Engineering/Biotechnology Biomedicine Biotechnology Cell Separation Diverse techniques Enzyme Activation Flow Cytometry Fluorescent Dyes Fundamental and applied biological sciences. Psychology Genes, Reporter Genome Humans Jurkat Cells Life Sciences Molecular and cellular biology research-article Signal Transduction Tetradecanoylphorbol Acetate - pharmacology Thapsigargin - pharmacology |
| title | A genome-wide functional assay of signal transduction in living mammalian cells |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-22T18%3A55%3A28IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20genome-wide%20functional%20assay%20of%20signal%20transduction%20in%20living%20mammalian%20cells&rft.jtitle=Nature%20biotechnology&rft.au=Whitney,%20Mike&rft.date=1998-12-01&rft.volume=16&rft.issue=13&rft.spage=1329&rft.epage=1333&rft.pages=1329-1333&rft.issn=1087-0156&rft.eissn=1546-1696&rft.coden=NABIF9&rft_id=info:doi/10.1038/4302&rft_dat=%3Cproquest_cross%3E17119285%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=17119285&rft_id=info:pmid/9853613&rfr_iscdi=true |