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Single cell RNA-sequencing on E16.5 and E17.5 mouse lungs

ABSTRACT: Single cell RNA-sequencing on E16.5 and E17.5 mouse lungs

PROVIDER: PRJNA657548 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12461511.fastq.gz	Fastqsanger.gz
	SRR12461512.fastq.gz	Fastqsanger.gz
	SRR12461513.fastq.gz	Fastqsanger.gz
	SRR12461514.fastq.gz	Fastqsanger.gz
	SRR12461515.fastq.gz	Fastqsanger.gz

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Project description:In this experiment we inhibited FGF signalling in E16.5 mouse lungs by using a dominant negative mouse model. We are interested in assessing the genetic regulation by FGF signalling at this stage of lung development. Specifically, we used a tetracycline controlled transcriptional activation model (Tet(o)) to manipulate FGF signalling. Genetically modified male mice that expressed a reverse tetracycline-controlled trans activator (rtTA) on the Rosa26 locus and a soluble FGFR2b on the Tet(o) promoter (Tet(o)sFgfr2b) (B6.Cg-Gt(ROSA)26Sortm1.1(rtTA,EGFP)NagyTg(tetO-Fgfr2b/Igh)1.3Jaw/sbel), were crossed with females lacking the Tet(o)sFgfr2b allele. Pregnant females were injected with doxycycline intraperitonally at E16.5. Doxycycline then induced the transcription soluble FGFR2b in the embryos, which inhibited FGF signalling by sequestering FGF ligands, preventing them from binding to the native FGFR2b receptor. We harvested embryonic lungs after 9 hrs of inhibition. We took the left lobe of each sample for RNA isolation. We set-up our matings so that 50% of the embryos in a litter were expected to contain the soluble FGFR2b. The remaining littermates will serve as the control group. We will compare the gene expression profiles of the experimental embryonic lungs with those of littermate controls.

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