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Using Next Generation Solexa Sequencing to Identify Genes that Regulate Stem Cell Proliferation in the Caenorhabditis elegans Germline
Author(s) -
Jones Vovanti,
Fox Paul,
Mardis Elaine,
Schedl Tim
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.699.1
Subject(s) - biology , germline , caenorhabditis elegans , genetics , rna interference , gene , phenocopy , gene silencing , mutant , rna
Multicellular organisms depend on cell‐cell interactions to coordinate their development. In C. elegans, germline stem cell fate depends on the activity of the Notch receptor homolog, glp‐1 . The loss of glp‐1 function prevents germ cell proliferation and produces sterile hermaphrodites. To identify new genes that regulate germline proliferation, suppressors of the temperature sensitive allele glp‐1(q231) have been isolated. These mutants, sog ( s uppressor o f g lp‐1 ), rescue sterility of q231 at 20°C. The sog‐1 gene was previously mapped to a 2 Mbp region on chrI. To [LL1]identify the gene responsible for the sog‐1 phenotype, we used Solexa sequencing to re‐sequence the entire genome of two independently isolated sog‐1 mutants. In the critical region, we identified 62 SNPs between the two strains and the reference genome. We performed RNAi screens of candidate genes to test whether they phenocopy sog‐1 suppression of glp‐1 . RNAi screens of pbrm‐1 displayed an embryonic lethal phenotype for all strains, while unc‐14 and F57B10.3 potentially phenocopy the germline suppression of q231 by sog‐1 . [LL2]Screens of other potential candidates were negative . These results may help identify new genes in germline proliferation and determine if Solexa sequencing will be useful in identifying genes defined by mutation. This work was funded by NIH/HIGMS MARC U*STAR T34 08663 National Research Service Award to UMBC, HHMI and BioMedRAP.