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Next Generation Sequencing Analysis of SRSF1 flox/flox and SRSF1-KO pancreatic islet Transcriptome

ABSTRACT: Next Generation Sequencing Analysis of SRSF1 flox/flox and SRSF1-KO pancreatic islet Transcriptome

PROVIDER: PRJNA725671 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14339947_1.fastq.gz	Fastqsanger.gz
	SRR14339947_2.fastq.gz	Fastqsanger.gz
	SRR14339948_1.fastq.gz	Fastqsanger.gz
	SRR14339948_2.fastq.gz	Fastqsanger.gz

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Next Generation Sequencing Analysis of SRSF1 flox/flox and SRSF1-KO pancreatic islet Transcriptome

Project description:we report insulin secretion defects in SRSF1 Ins2-Cre mice, in which SRSF1 is ablated in pancreatic β cells.

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MeRIP-Seq of pancreatic islets in Mettl3flox/flox and islet β cell-specific Mettl3 knockout (β-Mettl3-KO) mice

Project description:Purpose: Through mRNA m6A-profiling, we aim to characterize the mRNA m6A changes in pancreatic islets obtained from Mettl3flox/flox and β-Mettl3-KO mice. Methods: Total RNA was extracted using Tripure Isolation Reagent (Roche, Mannheim, Germany) from pancreatic islets of Mettl3flox/flox and β-Mettl3-KO mice at 8 weeks old. A total of 2-3 μg RNAs were pooled from nine Mettl3flox/flox mice and twelve β-Mettl3-KO mice, respectively. Three independent biological replicates of each group were used for MeRIP-seq. Fragmented RNA (~100 nt) was incubated for 2 hr at 4oC with anti-m6A polyclonal antibody (Merk Millipore) in the immunoprecipitation experiment. Then, immunoprecipitated RNAs or Input was used for library construction with Ovation SoLo RNA-Seq System Core Kit (NuGEN). The library preparations were sequenced on an Illumina Novaseq platform with a paired-end read length of 150 bp according to the standard protocols. After mapping reads to the reference genome, exomePeak R package (version 2.16.0) was used for the m6A peak identification in each anti-m6A immunoprecipitation group with the corresponding input samples serving as a control, and q-value threshold of enrichment of 0.05 was used for all data sets. The m6A-enriched motifs of each group were identified by HOMER (version 4.9.1). Conclusion: The islet mRNA m6A profiles in Mettl3flox/flox and β-Mettl3-KO mice were characterized.

2020-11-13 | GSE155614 | GEO

Next Generation Sequencing Analysis of Wild Type and SRSF1-MKO Skeletal Muscle Transcriptomes

Project description:Skeletal muscle specific SRSF1 KO mice were generated by mating SRSF1flox/flox and MyoD-Cre mice, Total RNA from muscle sample were analyzed by Next Generation Sequencing

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Project description:RNA-seq analysis was performed using pancreatic islet samples from flox mice and beta-cell specific CtBP2 KO mice.

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Project description:Next-generation sequencing analysis of 7-month-old SRSF2 flox/flox and SRSF2-PKO mice pancreatic islet transcriptome

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Brown Fat Tissue from Nipsnap1 Flox and Nipsnap1 KO mice via LC MS/MS

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MeRIP-Seq of pancreatic islets in Mettl3flox/flox and islet β cell-specific Mettl3 knockout (β-Mettl3-KO) mice

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Project description:Purpose: The goal of this study is to investigate how METTL3 regulates islet β-cell function. Methods: Total RNA was extracted using Tripure Isolation Reagent (Roche, Mannheim, Germany) from pancreatic islets of Mettl3flox/flox and β-Mettl3-KO mice at 8 weeks old. Each RNA sample was pooled from four Mettl3flox/flox and β-Mettl3-KO mice, respectively. Three independent biological replicates for each group were used for RNA-seq. RNA-seq was performed by deep sequencing using an Illumina Novaseq 6000 platform. Paired-end clean reads were aligned to the mouse reference genome(GRCm38.p6) with Hisat2 v2.0.5, and the aligned reads were used to quantify mRNA expression by using featureCounts v1.5.0-p3. Conclusion: Our study represents the first detailed analysis of islet transcriptomes from Mettl3flox/flox and β-Mettl3-KO mice, generated by RNA-seq technology. The RNA-seq analysis showed that 2560 genes were downregulated and 3408 genes were upregulated in the pancreatic islets of β-Mettl3-KO mice. GO analysis showed that the downregulated genes were primarily related to insulin secretion, SNARE binding, and mitochondrial respiratory chain, whereas the upregulated genes were associated with the immune response, B cell activation, and antigen binding.

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Project description:Mechanisms governing memory responses in IL-17 secreting CD4+ T cells (TH17), especially in autoimmune disorders, remain poorly understood. TH17 cells play pleiotropic roles in autoimmunity and tissue inflammation and are characterized by inherent plasticity, although how plasticity is regulated remains elusive. We investigated the transcriptional profiles of fate mapped (IL17aiCre-flox/stop/flox-YFP) TH17 cells and WT and Raptor-ko (flox/flox) TH17 cells (both IL17aiCre-flox/stop/flox-YFP) as well as sorted within CD27+ and CD27- subsets of WT in experimental autoimmune encephalomyelitis, a human model for human multiple sclerosis. We used microarrays to compare the global transcription profiles of YFP+ (IL17aiCre-flox/stop/flox-YFP) fate mapped TH17 cells as well as CD27+ and CD27- subsets of CD44+ fate mapped TH17 cells from draining lymph nodes on day 9 after immunization with Myelin oligodendrocyte glycoprotein from WT and IL17aCre-Raptor-flox/flox mice.

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