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MRNA sequencing of samples coming from Translating Ribosome Affinity Purification (TRAP) using MSH1P::FLAG::RPL18

ABSTRACT: mRNA sequencing of samples coming from Translating Ribosome Affinity Purification (TRAP) using MSH1P::FLAG::RPL18

PROVIDER: PRJNA470540 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR7135110_1.fastq.gz	Fastqsanger.gz
	SRR7135110_2.fastq.gz	Fastqsanger.gz
	SRR7135111_1.fastq.gz	Fastqsanger.gz
	SRR7135111_2.fastq.gz	Fastqsanger.gz
	SRR7135112_1.fastq.gz	Fastqsanger.gz

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mRNA sequencing of samples coming from Translating Ribosome Affinity Purification (TRAP) using MSH1P::FLAG::RPL18

Project description:Col-0 transgenic plants expressing MSH1P::FLAG::RPL18 were generated in both msh1 homozygous mutant (SAIL-877-F01) and wild type backgrounds with cloned MSH1 sequence described in Xu et al. (2011) and with transformation by the floral dip method (Clough and Bent, 1998). Positive transgenic lines were screened for FLAG tag by immunoblotting methods. Polysomal mRNA from MSH1-specific cells was obtained by ribosome purification using the TRAP method essentially as reported by (Reynoso et al., 2015) from floral stem tissues.

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Project description:We characterize a Cre-activated TRAP allele (Rosa26fsTRAP) to show that endothelium-specific activation of Rosa26fsTRAP identifies endothelial cell enriched transcripts, and that cardiomyocyte-restricted TRAP is a useful means to identify genes that are differentially expressed in cardiomyocytes in a disease model. We also show that TRAP is an effective means to study translational regulation, and we several nuclear-encoded mitochondrial genes are under strong translational control. These RNA-seq data show that the TRAP strategy and the Rosa26fsTRAP allele will be useful tools to probe cell type specific transcriptomes, and to study translational regulation. Study TRAP RNA and total RNA in two cases: heart specific tissues, Banding or Sham, using RNA-seq technology.

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