New Tools for Investigating the Comparative Biology of Caenorhabditis briggsae and C. elegans
Comparative studies of Caenorhabditis briggsae and C. elegans have provided insights into gene function and developmental control in both organisms. C. elegans is a well developed model organism with a variety of molecular and genetic tools to study gene functions. In contrast, there are only very l...
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| Veröffentlicht in: | Genetics (Austin) 2010-03, Vol.184 (3), p.853-863 |
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| creator | Zhao, Zhongying Flibotte, Stephane Murray, John I Blick, Daniel Boyle, Thomas J Gupta, Bhagwati Moerman, Donald G Waterston, Robert H |
| description | Comparative studies of Caenorhabditis briggsae and C. elegans have provided insights into gene function and developmental control in both organisms. C. elegans is a well developed model organism with a variety of molecular and genetic tools to study gene functions. In contrast, there are only very limited tools available for its closest relative, C. briggsae. To take advantage of the full potential of this comparative approach, we have developed several genetic and molecular tools to facilitate functional analysis in C. briggsae. First, we designed and implemented an SNP-based oligonucleotide microarray for rapid mapping of genetic mutants in C. briggsae. Second, we generated a mutagenized frozen library to permit the isolation of targeted deletions and used the library to recover a deletion mutant of cbr-unc-119 for use as a transgenic marker. Third, we used the cbr-unc-119 mutant in ballistic transformation and generated fluorescently labeled strains that allow automated lineaging and cellular resolution expression analysis. Finally, we demonstrated the potential of automated lineaging by profiling expression of egl-5, hlh-1, and pha-4 at cellular resolution and by detailed phenotyping of the perturbations on the Wnt signaling pathway. These additions to the experimental toolkit for C. briggsae should greatly increase its utility in comparative studies with C. elegans. With the emerging sequence of nematode species more closely related to C. briggsae, these tools may open novel avenues of experimentation in C. briggsae itself. |
| doi_str_mv | 10.1534/genetics.109.110270 |
| format | Article |
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C. elegans is a well developed model organism with a variety of molecular and genetic tools to study gene functions. In contrast, there are only very limited tools available for its closest relative, C. briggsae. To take advantage of the full potential of this comparative approach, we have developed several genetic and molecular tools to facilitate functional analysis in C. briggsae. First, we designed and implemented an SNP-based oligonucleotide microarray for rapid mapping of genetic mutants in C. briggsae. Second, we generated a mutagenized frozen library to permit the isolation of targeted deletions and used the library to recover a deletion mutant of cbr-unc-119 for use as a transgenic marker. Third, we used the cbr-unc-119 mutant in ballistic transformation and generated fluorescently labeled strains that allow automated lineaging and cellular resolution expression analysis. Finally, we demonstrated the potential of automated lineaging by profiling expression of egl-5, hlh-1, and pha-4 at cellular resolution and by detailed phenotyping of the perturbations on the Wnt signaling pathway. These additions to the experimental toolkit for C. briggsae should greatly increase its utility in comparative studies with C. elegans. With the emerging sequence of nematode species more closely related to C. briggsae, these tools may open novel avenues of experimentation in C. briggsae itself.</description><identifier>ISSN: 0016-6731</identifier><identifier>ISSN: 1943-2631</identifier><identifier>EISSN: 1943-2631</identifier><identifier>DOI: 10.1534/genetics.109.110270</identifier><identifier>PMID: 20008572</identifier><identifier>CODEN: GENTAE</identifier><language>eng</language><publisher>United States: Genetics Soc America</publisher><subject>Animals ; Binding sites ; Caenorhabditis elegans - genetics ; Caenorhabditis elegans - metabolism ; Chromosomes ; Comparative analysis ; Gene Expression Profiling - methods ; Gene Expression Regulation - physiology ; Genes, Helminth - physiology ; Genetic engineering ; Genomics ; Investigations ; Methods ; Models, Genetic ; Mutagenesis ; Nematodes ; Oligonucleotide Array Sequence Analysis ; Polymorphism, Single Nucleotide ; Sequence Deletion ; Studies</subject><ispartof>Genetics (Austin), 2010-03, Vol.184 (3), p.853-863</ispartof><rights>Copyright Genetics Society of America Mar 2010</rights><rights>Copyright © 2010 by the Genetics Society of America 2010</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c462t-5d0964c79fe097f00b71ce499b18c42d6eb5e58576ca85b3e8e6ce0ce2be22f53</citedby><cites>FETCH-LOGICAL-c462t-5d0964c79fe097f00b71ce499b18c42d6eb5e58576ca85b3e8e6ce0ce2be22f53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20008572$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhao, Zhongying</creatorcontrib><creatorcontrib>Flibotte, Stephane</creatorcontrib><creatorcontrib>Murray, John I</creatorcontrib><creatorcontrib>Blick, Daniel</creatorcontrib><creatorcontrib>Boyle, Thomas J</creatorcontrib><creatorcontrib>Gupta, Bhagwati</creatorcontrib><creatorcontrib>Moerman, Donald G</creatorcontrib><creatorcontrib>Waterston, Robert H</creatorcontrib><title>New Tools for Investigating the Comparative Biology of Caenorhabditis briggsae and C. elegans</title><title>Genetics (Austin)</title><addtitle>Genetics</addtitle><description>Comparative studies of Caenorhabditis briggsae and C. elegans have provided insights into gene function and developmental control in both organisms. C. elegans is a well developed model organism with a variety of molecular and genetic tools to study gene functions. In contrast, there are only very limited tools available for its closest relative, C. briggsae. To take advantage of the full potential of this comparative approach, we have developed several genetic and molecular tools to facilitate functional analysis in C. briggsae. First, we designed and implemented an SNP-based oligonucleotide microarray for rapid mapping of genetic mutants in C. briggsae. Second, we generated a mutagenized frozen library to permit the isolation of targeted deletions and used the library to recover a deletion mutant of cbr-unc-119 for use as a transgenic marker. Third, we used the cbr-unc-119 mutant in ballistic transformation and generated fluorescently labeled strains that allow automated lineaging and cellular resolution expression analysis. Finally, we demonstrated the potential of automated lineaging by profiling expression of egl-5, hlh-1, and pha-4 at cellular resolution and by detailed phenotyping of the perturbations on the Wnt signaling pathway. These additions to the experimental toolkit for C. briggsae should greatly increase its utility in comparative studies with C. elegans. With the emerging sequence of nematode species more closely related to C. briggsae, these tools may open novel avenues of experimentation in C. briggsae itself.</description><subject>Animals</subject><subject>Binding sites</subject><subject>Caenorhabditis elegans - genetics</subject><subject>Caenorhabditis elegans - metabolism</subject><subject>Chromosomes</subject><subject>Comparative analysis</subject><subject>Gene Expression Profiling - methods</subject><subject>Gene Expression Regulation - physiology</subject><subject>Genes, Helminth - physiology</subject><subject>Genetic engineering</subject><subject>Genomics</subject><subject>Investigations</subject><subject>Methods</subject><subject>Models, Genetic</subject><subject>Mutagenesis</subject><subject>Nematodes</subject><subject>Oligonucleotide Array Sequence Analysis</subject><subject>Polymorphism, Single Nucleotide</subject><subject>Sequence 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Finally, we demonstrated the potential of automated lineaging by profiling expression of egl-5, hlh-1, and pha-4 at cellular resolution and by detailed phenotyping of the perturbations on the Wnt signaling pathway. These additions to the experimental toolkit for C. briggsae should greatly increase its utility in comparative studies with C. elegans. With the emerging sequence of nematode species more closely related to C. briggsae, these tools may open novel avenues of experimentation in C. briggsae itself.</abstract><cop>United States</cop><pub>Genetics Soc America</pub><pmid>20008572</pmid><doi>10.1534/genetics.109.110270</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Binding sites Caenorhabditis elegans - genetics Caenorhabditis elegans - metabolism Chromosomes Comparative analysis Gene Expression Profiling - methods Gene Expression Regulation - physiology Genes, Helminth - physiology Genetic engineering Genomics Investigations Methods Models, Genetic Mutagenesis Nematodes Oligonucleotide Array Sequence Analysis Polymorphism, Single Nucleotide Sequence Deletion Studies |
| title | New Tools for Investigating the Comparative Biology of Caenorhabditis briggsae and C. elegans |
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