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M6A methylation regulates the fate of endogenous retrovirus transcripts [RIP-seq]

ABSTRACT: m6A methylation regulates the fate of endogenous retrovirus transcripts [RIP-seq]

PROVIDER: PRJNA606743 | ENA |

REPOSITORIES: ENA

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Dataset's files

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			Action	DRS
	SRR11088735_1.fastq.gz	Fastqsanger.gz
	SRR11088735_2.fastq.gz	Fastqsanger.gz
	SRR11088736_1.fastq.gz	Fastqsanger.gz
	SRR11088736_2.fastq.gz	Fastqsanger.gz
	SRR11088737_1.fastq.gz	Fastqsanger.gz

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m6A methylation regulates the fate of endogenous retrovirus transcripts [RIP-seq]

Project description:Using a genome-scale CRISPR knockout screening in mouse embryonic stem cells, we identify m6A RNA methylation as an important regulatory component that restricts the activity of endogenous retroviruses of the IAP (intracisternal A-particles) family. The m6A methylation of IAP mRNAs occurs on their 5’ end, is catalyzed by the complex of methyltransferase-like (METTL)3/METTL14 proteins whose depletion, along with other complex subunits, namely WTAP and ZC3H13, leads to increased IAP transcript abundance. Using auxin-dependent degron, we show that rapid removal of METTL3, METTL14 and ZC3H13 further increases IAP RNA abundance in a progressive and reversible fashion. Finally, we demonstrate that m6A RNA reduces the stability of IAP transcripts

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Project description:We performed m6A-RIPs in Ascl1-induced neurons (iNeurons) to investigate the neuronal m6A epitranscriptome. Immunoprecipitation was done twice using two different antibodies, acquired from Abcam and Synaptic Systems (SySy), allowing for a more robust detection of m6A modification marks. Additionally, RIP-seq was performed separately with intact and fragmented RNA. The former approach allowed to identify proportions of m6A-modified transcripts among the total number, while the latter approach provided the information to identify genomic coordinates of m6A peaks.

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