| Budget Amount *help |
¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2014: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2012: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
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| Outline of Final Research Achievements |
Inhibition of GPI-anchored protein (GPI-AP) synthesis of C. elegans results in severe germline abnormalities. By analyzing a deletion allele of the GPI-anchor synthesizing gene piga-1, we confirmed that the phenotypes were caused by the decrease of dividing germline stem cells (GSCs) at the distal end of the gonad where distal tip cell is providing Notch signal to maintain GSC niche. In this study, we studied GPI-APs enriched in the germline by biochemical analysis, by DNA microarray and RNA-Seq. We also examined RNAi phenotypes of all the GPI-AP genes to identify essential GPI-APs in the GSC niche. From these analyses, we identified seven different GPI-AP coding genes. Among them, two genes seem to be especially important. Thus, we isolated CRISPR-Cas9 mediated deletion allele of the two genes, and found that the germline formation was severely affected in the allele. We are examining the possible involvement of these two genes in the Notch signaling pathways of the GSCs.
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