An antisense-based functional genomics approach for identification of genes critical for growth of Candida albicans
Converting the complete genome sequence of Candida albicans into meaningful biological information will require comprehensive screens for identifying functional classes of genes. Most systems described so far are not applicable to C. albicans because of its difficulty with mating, its diploid nature...
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| Veröffentlicht in: | Nature biotechnology 2001-03, Vol.19 (3), p.235-241 |
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| creator | De Backer, Marianne D. Nelissen, Bart Logghe, Marc Viaene, Jasmine Loonen, Inge Vandoninck, Sandy de Hoogt, Ronald Dewaele, Sylviane Simons, Fermin A. Verhasselt, Peter Vanhoof, Greet Contreras, Roland Luyten, Walter H.M.L. |
| description | Converting the complete genome sequence of
Candida albicans
into meaningful biological information will require comprehensive screens for identifying functional classes of genes. Most systems described so far are not applicable to
C. albicans
because of its difficulty with mating, its diploid nature, and the lack of functional random insertional mutagenesis methods. We examined artificial gene suppression as a means to identify gene products critical for growth of this pathogen; these represent new antifungal drug targets. To achieve gene suppression we combined antisense RNA inhibition and promoter interference. After cloning antisense complementary DNA (cDNA) fragments under control of an inducible GAL1 promoter, we transferred the resulting libraries to
C. albicans
. Over 2,000 transformant colonies were screened for a promoter-induced diminished-growth phenotype. After recovery of the plasmids, sequence determination of their inserts revealed the messenger RNA (mRNA) they inhibited or the gene they disrupted. Eighty-six genes critical for growth were identified, 45 with unknown function. When used in high-throughput screening for antifungals, the crippled
C. albicans
strains generated in this study showed enhanced sensitivity to specific drugs. |
| doi_str_mv | 10.1038/85677 |
| format | Article |
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Candida albicans
into meaningful biological information will require comprehensive screens for identifying functional classes of genes. Most systems described so far are not applicable to
C. albicans
because of its difficulty with mating, its diploid nature, and the lack of functional random insertional mutagenesis methods. We examined artificial gene suppression as a means to identify gene products critical for growth of this pathogen; these represent new antifungal drug targets. To achieve gene suppression we combined antisense RNA inhibition and promoter interference. After cloning antisense complementary DNA (cDNA) fragments under control of an inducible GAL1 promoter, we transferred the resulting libraries to
C. albicans
. Over 2,000 transformant colonies were screened for a promoter-induced diminished-growth phenotype. After recovery of the plasmids, sequence determination of their inserts revealed the messenger RNA (mRNA) they inhibited or the gene they disrupted. Eighty-six genes critical for growth were identified, 45 with unknown function. When used in high-throughput screening for antifungals, the crippled
C. albicans
strains generated in this study showed enhanced sensitivity to specific drugs.</description><identifier>ISSN: 1087-0156</identifier><identifier>EISSN: 1546-1696</identifier><identifier>DOI: 10.1038/85677</identifier><identifier>PMID: 11231556</identifier><identifier>CODEN: NABIF9</identifier><language>eng</language><publisher>New York: Nature Publishing Group US</publisher><subject>Agriculture ; Antibiotics ; Antifungal Agents - pharmacology ; Bioinformatics ; Biological and medical sciences ; Biomedical and Life Sciences ; Biomedical Engineering/Biotechnology ; Biomedicine ; Biotechnology ; Candida albicans ; Candida albicans - drug effects ; Candida albicans - genetics ; Candida albicans - growth & development ; Cloning ; Cloning, Molecular - methods ; Deoxyribonucleic acid ; Diverse techniques ; DNA ; DNA, Antisense - genetics ; Drug Evaluation, Preclinical ; Fundamental and applied biological sciences. Psychology ; Fungal Proteins - genetics ; Fungal Proteins - metabolism ; Gene Expression Regulation, Fungal ; Gene Library ; Genes, Essential - genetics ; Genes, Fungal - genetics ; Genome, Fungal ; genomics ; Genomics - methods ; Health. Pharmaceutical industry ; Heterozygote ; high-throughput screening ; Industrial applications and implications. Economical aspects ; Life Sciences ; Microbial Sensitivity Tests ; Molecular and cellular biology ; Mutagenesis, Insertional - genetics ; Pathogens ; Phenotype ; Production of active biomolecules ; Promoter Regions, Genetic - genetics ; RNA, Antisense - genetics ; RNA, Fungal - analysis ; RNA, Fungal - genetics ; RNA, Messenger - analysis ; RNA, Messenger - genetics ; Transformation, Genetic</subject><ispartof>Nature biotechnology, 2001-03, Vol.19 (3), p.235-241</ispartof><rights>Springer Nature America, Inc. 2001</rights><rights>2001 INIST-CNRS</rights><rights>COPYRIGHT 2001 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group Mar 2001</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c601t-89c5b9780d22e66200317067b7d3d69511b0bea0c9182ace264a75253b0528e63</citedby><cites>FETCH-LOGICAL-c601t-89c5b9780d22e66200317067b7d3d69511b0bea0c9182ace264a75253b0528e63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/85677$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/85677$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1034250$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11231556$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>De Backer, Marianne D.</creatorcontrib><creatorcontrib>Nelissen, Bart</creatorcontrib><creatorcontrib>Logghe, Marc</creatorcontrib><creatorcontrib>Viaene, Jasmine</creatorcontrib><creatorcontrib>Loonen, Inge</creatorcontrib><creatorcontrib>Vandoninck, Sandy</creatorcontrib><creatorcontrib>de Hoogt, Ronald</creatorcontrib><creatorcontrib>Dewaele, Sylviane</creatorcontrib><creatorcontrib>Simons, Fermin A.</creatorcontrib><creatorcontrib>Verhasselt, Peter</creatorcontrib><creatorcontrib>Vanhoof, Greet</creatorcontrib><creatorcontrib>Contreras, Roland</creatorcontrib><creatorcontrib>Luyten, Walter H.M.L.</creatorcontrib><title>An antisense-based functional genomics approach for identification of genes critical for growth of Candida albicans</title><title>Nature biotechnology</title><addtitle>Nat Biotechnol</addtitle><addtitle>Nat Biotechnol</addtitle><description>Converting the complete genome sequence of
Candida albicans
into meaningful biological information will require comprehensive screens for identifying functional classes of genes. Most systems described so far are not applicable to
C. albicans
because of its difficulty with mating, its diploid nature, and the lack of functional random insertional mutagenesis methods. We examined artificial gene suppression as a means to identify gene products critical for growth of this pathogen; these represent new antifungal drug targets. To achieve gene suppression we combined antisense RNA inhibition and promoter interference. After cloning antisense complementary DNA (cDNA) fragments under control of an inducible GAL1 promoter, we transferred the resulting libraries to
C. albicans
. Over 2,000 transformant colonies were screened for a promoter-induced diminished-growth phenotype. After recovery of the plasmids, sequence determination of their inserts revealed the messenger RNA (mRNA) they inhibited or the gene they disrupted. Eighty-six genes critical for growth were identified, 45 with unknown function. When used in high-throughput screening for antifungals, the crippled
C. albicans
strains generated in this study showed enhanced sensitivity to specific drugs.</description><subject>Agriculture</subject><subject>Antibiotics</subject><subject>Antifungal Agents - pharmacology</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>Candida albicans</subject><subject>Candida albicans - drug effects</subject><subject>Candida albicans - genetics</subject><subject>Candida albicans - growth & development</subject><subject>Cloning</subject><subject>Cloning, Molecular - methods</subject><subject>Deoxyribonucleic acid</subject><subject>Diverse techniques</subject><subject>DNA</subject><subject>DNA, Antisense - genetics</subject><subject>Drug Evaluation, Preclinical</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fungal Proteins - genetics</subject><subject>Fungal Proteins - metabolism</subject><subject>Gene Expression Regulation, Fungal</subject><subject>Gene Library</subject><subject>Genes, Essential - genetics</subject><subject>Genes, Fungal - genetics</subject><subject>Genome, Fungal</subject><subject>genomics</subject><subject>Genomics - methods</subject><subject>Health. Pharmaceutical industry</subject><subject>Heterozygote</subject><subject>high-throughput screening</subject><subject>Industrial applications and implications. Economical aspects</subject><subject>Life Sciences</subject><subject>Microbial Sensitivity Tests</subject><subject>Molecular and cellular biology</subject><subject>Mutagenesis, Insertional - genetics</subject><subject>Pathogens</subject><subject>Phenotype</subject><subject>Production of active biomolecules</subject><subject>Promoter Regions, Genetic - genetics</subject><subject>RNA, Antisense - genetics</subject><subject>RNA, Fungal - analysis</subject><subject>RNA, Fungal - genetics</subject><subject>RNA, Messenger - analysis</subject><subject>RNA, Messenger - genetics</subject><subject>Transformation, Genetic</subject><issn>1087-0156</issn><issn>1546-1696</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>BENPR</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqNkk1r3DAQhkVpaNI0f6GY0gZ6cCrJlmQflyVpA4FAv65mLI8dBa-01di0_ffRfsB2ewo6SMw8M6N5Zxi7EPxK8KL6VCltzAt2JlSpc6Fr_TK9eWVyLpQ-Za-JHjnnutT6FTsVQhZCKX3GaOEz8JMj9IR5C4Rd1s_eTi54GLMBfVg5Sxms1zGAfcj6EDPXYQrpnYUNloV-wyFlNropGcctNMTwe3rYOJfgO9dBBmObvJ7esJMeRsKL_X3Oftxcf19-ye_uP98uF3e51VxMeVVb1dam4p2UqLXkvBCGa9Oaruh0rYRoeYvAbS0qCRalLsEoqYqWK1mhLs7Z5S5v-vqvGWlqVo4sjiN4DDM1wlSlUEYl8N1_4GOYY-qfGillkbTSPEFXO2iAERvn-zBFsOl0mBQKHnuX7AtRJ5nrYhvw8SggMRP-mQaYiZrbb1-fz97_PGY_7FgbA1HEvllHt4L4txG82exCs92FxL3ddzW3K-wO1H74CXi_B4DS1PoI3jr6J1tRSsUPMlLy-AHjQZ3jgk-Tz8Oq</recordid><startdate>20010301</startdate><enddate>20010301</enddate><creator>De Backer, Marianne D.</creator><creator>Nelissen, Bart</creator><creator>Logghe, Marc</creator><creator>Viaene, Jasmine</creator><creator>Loonen, Inge</creator><creator>Vandoninck, Sandy</creator><creator>de Hoogt, Ronald</creator><creator>Dewaele, Sylviane</creator><creator>Simons, Fermin A.</creator><creator>Verhasselt, Peter</creator><creator>Vanhoof, Greet</creator><creator>Contreras, Roland</creator><creator>Luyten, Walter H.M.L.</creator><general>Nature Publishing Group US</general><general>Nature</general><general>Nature Publishing Group</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QO</scope><scope>7QP</scope><scope>7QR</scope><scope>7T7</scope><scope>7TK</scope><scope>7TM</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>M2P</scope><scope>M7P</scope><scope>M7S</scope><scope>MBDVC</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>Q9U</scope><scope>RC3</scope><scope>M7N</scope></search><sort><creationdate>20010301</creationdate><title>An antisense-based functional genomics approach for identification of genes critical for growth of Candida albicans</title><author>De Backer, Marianne D. ; Nelissen, Bart ; Logghe, Marc ; Viaene, Jasmine ; Loonen, Inge ; Vandoninck, Sandy ; de Hoogt, Ronald ; Dewaele, Sylviane ; Simons, Fermin A. ; Verhasselt, Peter ; Vanhoof, Greet ; Contreras, Roland ; Luyten, Walter H.M.L.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c601t-89c5b9780d22e66200317067b7d3d69511b0bea0c9182ace264a75253b0528e63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Agriculture</topic><topic>Antibiotics</topic><topic>Antifungal Agents - pharmacology</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>Candida albicans</topic><topic>Candida albicans - drug effects</topic><topic>Candida albicans - genetics</topic><topic>Candida albicans - growth & development</topic><topic>Cloning</topic><topic>Cloning, Molecular - methods</topic><topic>Deoxyribonucleic acid</topic><topic>Diverse techniques</topic><topic>DNA</topic><topic>DNA, Antisense - genetics</topic><topic>Drug Evaluation, Preclinical</topic><topic>Fundamental and applied biological sciences. 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complete genome sequence of
Candida albicans
into meaningful biological information will require comprehensive screens for identifying functional classes of genes. Most systems described so far are not applicable to
C. albicans
because of its difficulty with mating, its diploid nature, and the lack of functional random insertional mutagenesis methods. We examined artificial gene suppression as a means to identify gene products critical for growth of this pathogen; these represent new antifungal drug targets. To achieve gene suppression we combined antisense RNA inhibition and promoter interference. After cloning antisense complementary DNA (cDNA) fragments under control of an inducible GAL1 promoter, we transferred the resulting libraries to
C. albicans
. Over 2,000 transformant colonies were screened for a promoter-induced diminished-growth phenotype. After recovery of the plasmids, sequence determination of their inserts revealed the messenger RNA (mRNA) they inhibited or the gene they disrupted. Eighty-six genes critical for growth were identified, 45 with unknown function. When used in high-throughput screening for antifungals, the crippled
C. albicans
strains generated in this study showed enhanced sensitivity to specific drugs.</abstract><cop>New York</cop><pub>Nature Publishing Group US</pub><pmid>11231556</pmid><doi>10.1038/85677</doi><tpages>7</tpages></addata></record> |
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| ispartof | Nature biotechnology, 2001-03, Vol.19 (3), p.235-241 |
| issn | 1087-0156 1546-1696 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_17841575 |
| source | MEDLINE; Springer Nature - Complete Springer Journals; Nature Journals Online |
| subjects | Agriculture Antibiotics Antifungal Agents - pharmacology Bioinformatics Biological and medical sciences Biomedical and Life Sciences Biomedical Engineering/Biotechnology Biomedicine Biotechnology Candida albicans Candida albicans - drug effects Candida albicans - genetics Candida albicans - growth & development Cloning Cloning, Molecular - methods Deoxyribonucleic acid Diverse techniques DNA DNA, Antisense - genetics Drug Evaluation, Preclinical Fundamental and applied biological sciences. Psychology Fungal Proteins - genetics Fungal Proteins - metabolism Gene Expression Regulation, Fungal Gene Library Genes, Essential - genetics Genes, Fungal - genetics Genome, Fungal genomics Genomics - methods Health. Pharmaceutical industry Heterozygote high-throughput screening Industrial applications and implications. Economical aspects Life Sciences Microbial Sensitivity Tests Molecular and cellular biology Mutagenesis, Insertional - genetics Pathogens Phenotype Production of active biomolecules Promoter Regions, Genetic - genetics RNA, Antisense - genetics RNA, Fungal - analysis RNA, Fungal - genetics RNA, Messenger - analysis RNA, Messenger - genetics Transformation, Genetic |
| title | An antisense-based functional genomics approach for identification of genes critical for growth of Candida albicans |
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