E-MTAB-7063
RNA-Seq  to identify germline stem cell-enriched genes and cystoblast-enriched genes and BMP targeted genes
We sequenced the transcriptomes of genetically expanded stem cells that either experience high levels of BMP signalling (expression of constitutively active BMPR, tkvQD.) or low levels (expression of shRNA again the differentiation promoting factor bam). We also sequenced RNA from differentiating daughter cells (cystoblasts) isolated through the expression of bam.GFP. We then performed a differential expression analysis. Our aim was to identify germline stem cell-enriched gene and cystoblast-enriched genes by tkvQD vs bam-GFP. We also wished to identify possible BMP target genes by comparing tkvQD and bam.shRNA. We have subsequently taken a few of the genes identified and validated their roles in germline stem cell self-renewal and differentiation in vivo.
We sequenced the transcriptomes of genetically expanded stem cells that either experience high levels of BMP signalling (expression of constitutively active BMPR, tkvQD.) or low levels (expression of shRNA again the differentiation promoting factor bam). We also sequenced RNA from differentiating daughter cells (cystoblasts) isolated through the expression of bam.GFP. We then performed a differential expression analysis. Our aim was to identify germline stem cell-enriched gene and cystoblast-enriched genes by tkvQD vs bam-GFP. We also wished to identify possible BMP target genes by comparing tkvQD and bam.shRNA. We have subsequently taken a few of the genes identified and validated their roles in germline stem cell self-renewal and differentiation in vivo.