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Small RNA sequencing of spermatogonia with different m6a methylation

ABSTRACT: small RNA sequencing of spermatogonia with different m6a methylation

PROVIDER: PRJNA833869 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR19023387.fastq.gz	Fastqsanger.gz
	SRR19023388.fastq.gz	Fastqsanger.gz
	SRR19023389.fastq.gz	Fastqsanger.gz
	SRR19023390.fastq.gz	Fastqsanger.gz
	SRR19023391.fastq.gz	Fastqsanger.gz

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small RNA sequencing of spermatogonia with different m6a methylation

Project description:we knocked down the expression level of METTL3 in spermatogonia to reduce the total m6A methylation in the cells, and mapped the small RNA expression profile of spermatogonia after the total m6A methylation level was reduced，and we after the m6A methylation was reduced, many miRNA and circRNA were differently expressed

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RNA sequencing of spermatogonia with different m6a methylation

Project description:RNA sequencing of spermatogonia with different m6a methylation

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m6A RNA methylation promotes XIST-mediated transcriptional repression

Project description:XIST is a long non-coding RNA (lncRNA) that mediates transcriptional silencing of X chromosome genes. Here we show that XIST is highly methylated with at least 78 N6-methyladenosine (m6A) residues, a reversible base modification whose function in lncRNAs is unknown. We show that m6A formation in XIST, as well as cellular mRNAs, is mediated by RBM15 and its paralog RBM15B, which bind the m6A-methylation complex and recruit it to specific sites in RNA. This results in methylation of adenosines in adjacent m6A consensus motifs. Furthermore, knockdown of RBM15 and RBM15B, or knockdown of the m6A methyltransferase METTL3 impairs XIST-mediated gene silencing. A systematic comparison of m6A-binding proteins shows that YTHDC1 preferentially recognizes m6A in XIST and is required for XIST function. Additionally, artificial tethering of YTHDC1 to XIST rescues XIST-mediated silencing upon loss of m6A. These data reveal a pathway of m6A formation and recognition required for XIST-mediated transcriptional repression. Three to four biological HEK293T replicates were used to perform iCLIP of endogenous YTH proteins, RBM15, and RBM15B. Crosslinking induced truncations were identified using CIMS-CITS pipeline.

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Project description:Spermatogenesis, an efficient and complex system in male germline development, requires a series of elaborately regulated genetic events in which diploid spermatogonia differentiate into haploid spermatozoa. N6-methyladenosine (m6A) is important for spermatogenesis. ALKBH5 is an m6A eraser, and knockout of ALKBH5 increases the level of total m6A methylation and causes male infertility.

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