Identification of functional genes during Fas-mediated apoptosis using a randomly fragmented cDNA library
We describe a general strategy for the identification of functional genes that, when downregulated, result in a selectable phenotype. This strategy is based on expression selection of cDNA fragments that counteract their cognate genes. A cDNA library containing random fragments expressed in human He...
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| Veröffentlicht in: | Cellular and molecular life sciences : CMLS 2005-09, Vol.62 (17), p.2015-2026 |
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| container_title | Cellular and molecular life sciences : CMLS |
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| creator | Hassan, M Mirmohammadsadegh, A Selimovic, D Nambiar, S Tannapfel, A Hengge, U R |
| description | We describe a general strategy for the identification of functional genes that, when downregulated, result in a selectable phenotype. This strategy is based on expression selection of cDNA fragments that counteract their cognate genes. A cDNA library containing random fragments expressed in human HepG2, A375 and CLS-354 cells was used to identify functional genes whose inhibition conferred resistance to Fas-induced apoptosis. Thirty-five clones were isolated, 28 of which were derived from unknown genes, that tagged 19 individual genes and 7 of which referred to known genes that tagged the apoptosis-related protein (APR)-1, -2 and indoleamine-pyrrole 2,3,-dioxygenase (IDO). The ability of APR-1-, -2- and IDO-derived antisense RNAs to induce resistance to Fas in HepG2, A375 and CLS-354 cells suggested that APR-1, -2 and IDO genes are involved in the machinery of Fas-mediated apoptosis. Our gene discovery strategy provides a generally applicable procedure to identify functional genes that interfere with apoptosis, and may therefore be clinically relevant for tumor therapy. |
| doi_str_mv | 10.1007/s00018-005-5172-6 |
| format | Article |
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This strategy is based on expression selection of cDNA fragments that counteract their cognate genes. A cDNA library containing random fragments expressed in human HepG2, A375 and CLS-354 cells was used to identify functional genes whose inhibition conferred resistance to Fas-induced apoptosis. Thirty-five clones were isolated, 28 of which were derived from unknown genes, that tagged 19 individual genes and 7 of which referred to known genes that tagged the apoptosis-related protein (APR)-1, -2 and indoleamine-pyrrole 2,3,-dioxygenase (IDO). The ability of APR-1-, -2- and IDO-derived antisense RNAs to induce resistance to Fas in HepG2, A375 and CLS-354 cells suggested that APR-1, -2 and IDO genes are involved in the machinery of Fas-mediated apoptosis. Our gene discovery strategy provides a generally applicable procedure to identify functional genes that interfere with apoptosis, and may therefore be clinically relevant for tumor therapy.</description><identifier>ISSN: 1420-682X</identifier><identifier>EISSN: 1420-9071</identifier><identifier>DOI: 10.1007/s00018-005-5172-6</identifier><identifier>PMID: 16091844</identifier><language>eng</language><publisher>Switzerland: Springer Nature B.V</publisher><subject>Antibodies - pharmacology ; Apoptosis ; Apoptosis - genetics ; Cells, Cultured ; Deoxyribonucleic acid ; DNA ; fas Receptor - drug effects ; fas Receptor - immunology ; Gene Expression ; Gene Library ; Genotype & phenotype ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; Interferon-gamma - pharmacology ; Phenotype ; RNA, Antisense - genetics ; RNA, Antisense - isolation & purification ; RNA, Antisense - metabolism ; Tryptophan Oxygenase - antagonists & inhibitors ; Tryptophan Oxygenase - genetics ; Tryptophan Oxygenase - metabolism ; Tumor Cells, Cultured</subject><ispartof>Cellular and molecular life sciences : CMLS, 2005-09, Vol.62 (17), p.2015-2026</ispartof><rights>Birkhäuser Verlag, Basel 2005</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c477t-7496ff762dce9d3c42d35a92cd4d3742efc069a70962e869406a05491f2d6c803</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC11139157/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC11139157/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16091844$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hassan, M</creatorcontrib><creatorcontrib>Mirmohammadsadegh, A</creatorcontrib><creatorcontrib>Selimovic, D</creatorcontrib><creatorcontrib>Nambiar, S</creatorcontrib><creatorcontrib>Tannapfel, A</creatorcontrib><creatorcontrib>Hengge, U R</creatorcontrib><title>Identification of functional genes during Fas-mediated apoptosis using a randomly fragmented cDNA library</title><title>Cellular and molecular life sciences : CMLS</title><addtitle>Cell Mol Life Sci</addtitle><description>We describe a general strategy for the identification of functional genes that, when downregulated, result in a selectable phenotype. This strategy is based on expression selection of cDNA fragments that counteract their cognate genes. A cDNA library containing random fragments expressed in human HepG2, A375 and CLS-354 cells was used to identify functional genes whose inhibition conferred resistance to Fas-induced apoptosis. Thirty-five clones were isolated, 28 of which were derived from unknown genes, that tagged 19 individual genes and 7 of which referred to known genes that tagged the apoptosis-related protein (APR)-1, -2 and indoleamine-pyrrole 2,3,-dioxygenase (IDO). The ability of APR-1-, -2- and IDO-derived antisense RNAs to induce resistance to Fas in HepG2, A375 and CLS-354 cells suggested that APR-1, -2 and IDO genes are involved in the machinery of Fas-mediated apoptosis. Our gene discovery strategy provides a generally applicable procedure to identify functional genes that interfere with apoptosis, and may therefore be clinically relevant for tumor therapy.</description><subject>Antibodies - pharmacology</subject><subject>Apoptosis</subject><subject>Apoptosis - genetics</subject><subject>Cells, Cultured</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>fas Receptor - drug effects</subject><subject>fas Receptor - immunology</subject><subject>Gene Expression</subject><subject>Gene Library</subject><subject>Genotype & phenotype</subject><subject>Humans</subject><subject>Indoleamine-Pyrrole 2,3,-Dioxygenase</subject><subject>Interferon-gamma - pharmacology</subject><subject>Phenotype</subject><subject>RNA, Antisense - genetics</subject><subject>RNA, Antisense - isolation & purification</subject><subject>RNA, Antisense - metabolism</subject><subject>Tryptophan Oxygenase - antagonists & inhibitors</subject><subject>Tryptophan Oxygenase - 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| subjects | Antibodies - pharmacology Apoptosis Apoptosis - genetics Cells, Cultured Deoxyribonucleic acid DNA fas Receptor - drug effects fas Receptor - immunology Gene Expression Gene Library Genotype & phenotype Humans Indoleamine-Pyrrole 2,3,-Dioxygenase Interferon-gamma - pharmacology Phenotype RNA, Antisense - genetics RNA, Antisense - isolation & purification RNA, Antisense - metabolism Tryptophan Oxygenase - antagonists & inhibitors Tryptophan Oxygenase - genetics Tryptophan Oxygenase - metabolism Tumor Cells, Cultured |
| title | Identification of functional genes during Fas-mediated apoptosis using a randomly fragmented cDNA library |
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