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Mettl3-RIP-sequencing of NK cells

ABSTRACT: Mettl3-RIP-sequencing of NK cells

PROVIDER: PRJNA725188 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14325930_1.fastq.gz	Fastqsanger.gz
	SRR14325930_2.fastq.gz	Fastqsanger.gz
	SRR14325931_1.fastq.gz	Fastqsanger.gz
	SRR14325931_2.fastq.gz	Fastqsanger.gz
	SRR14325932_1.fastq.gz	Fastqsanger.gz

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Mettl3-RIP-sequencing of NK cells

Project description:m6A modification plays vital roles in regulating mRNA lifecycle, thus controlling the biological process of multiple cell types. Here we intended to discover the binding sites of Mettl3 protein on mRNA of NK cells. Total RNAs were isolated from WT splenic NK cells, and subjected to standard RIP protocol. The library products corresponding to 200-500 bps were enriched, quantified and finally sequenced on Illumina Novaseq 6000 sequencer with PE150 model. Mettl3 binding sites were located primarily in the 3’ untranslated regions (UTRs), coding sequences (CDSs) region and near stop codons.

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Project description:Purpose: For comparing the transcript changes, we conducted mRNA-sequencing of splenic NK cells from Ncr1Cre-Mettl3fl/fl (cKO) and Mettl3fl/fl (WT) mice. Method: Firstly, The splenic NK cells (CD45.2+CD3-NK1.1+NKp46+) are purified via Fluorescence activated Cell Sorting (FACS), then frozen in -80 °C ultra-low temperature refrigerator, followed by High-throughput sequencing, in three replicates, using Illumina Hiseq 1500 platform. Result: Using standard data process workflow, we found the differentailly expressed genes in NK cells from Ncr1Cre-Mettl3fl/fl (cKO) mice, compared with those from Mettl3fl/fl (WT) mice.

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