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SRX2325626: GSM2367701: kmg KD testis rep2; Drosophila melanogaster; RNA-Seq
1 ILLUMINA (NextSeq 500) run: 46.8M spots, 14.1G bases, 6.1Gb downloads

Submitted by: NCBI (GEO)
Study: Gene expression of fly testes with dMi-2, kumgang (CG5204) knock downs and wild-type heads by RNA-Seq
show Abstracthide Abstract
The effect of different spermatocyte-specific loss of functions; kumgang (kmg or CG5204), dMi-2 in the gene expression in fly testes was assessed by RNA-Seq. Gene expression in wild-type heads were also measured to have a reference expression profile of ''somatic tissues''. / Title: Blocking promiscuous activation at cryptic promoters directs cell type–specific gene expression / Abstract: To selectively express cell type–specific transcripts during development, it is critical to maintain genes required for other lineages in a silent state. Here, we show in the Drosophila male germline stem cell lineage that a spermatocyte-specific zinc finger protein, Kumgang (Kmg), working with the chromatin remodeler dMi-2 prevents transcription of genes normally expressed only in somatic lineages. By blocking transcription from normally cryptic promoters, Kmg restricts activation by Aly, a component of the testis-meiotic arrest complex, to transcripts for male germ cell differentiation. Our results suggest that as new regions of the genome become open for transcription during terminal differentiation, blocking the action of a promiscuous activator on cryptic promoters is a critical mechanism for specifying precise gene activation. Overall design: Gene expression profile of kumgang and dMi-2 knock down testes and corresponding sibling control testes (no Gal4 driver), and wild-type heads were generated in duplicate using Illumina NextSeq 500.
Sample: kmg KD testis rep2
SAMN05978655 • SRS1781461 • All experiments • All runs
Library:
Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: kmg ctrl, kmg KD, dMi-2 ctrl, and dMi-2 KD testes without accessory glands were dissected in PBS for 30 minutes and immediately frozen in liquid nitrogen before transferred to -80°C. Wild-type heads were manually decapitated from the body and frozen and stored as in testes preparation. Total RNA was extracted using TRIzol (Invitrogen). RNA was further purified by RNeasy mini column (Qiagen) to minimize residual salts. RNA quality was assessed by checking rRNA integrity using Bioanalyzer. ~30ug of total RNA was used to purify polyA RNA by using Poly(A)Purist MAG Kit (Thermo Fisher, Cat#AM1922). 100ng of purified mRNA was used to make a strand-specific sequencing library by dUTP incorporation method using KAPA Stranded RNA-Seq Library Preparation Kit Illumina® Platforms (KAPA Biosystems, Cat#: KK8400) following manufacturer’s instructions. All RNA-Seq libraries were sequenced by NextSeq Mid Output mode 150bp paired end. Between 3 to 4 samples were pooled in a single lane, yielding on average 48 million reads per sample (34 to 65 million). All sequencing was performed by the Stanford Functional Genomics Facility (SFGF).
Experiment attributes:
GEO Accession: GSM2367701
Links:
Runs: 1 run, 46.8M spots, 14.1G bases, 6.1Gb
Run# of Spots# of BasesSizePublished
SRR489631446,840,80014.1G6.1Gb2017-05-19

ID:
3399496

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