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"TRADES": Targeted RNA demethylation by Suntag system

ABSTRACT: "TRADES": Targeted RNA demethylation by Suntag system

PROVIDER: PRJNA586772 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10377460_1.fastq.gz	Fastqsanger.gz
	SRR10377460_2.fastq.gz	Fastqsanger.gz
	SRR10377461_1.fastq.gz	Fastqsanger.gz
	SRR10377461_2.fastq.gz	Fastqsanger.gz
	SRR10377462_1.fastq.gz	Fastqsanger.gz

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Project description:DNA methylation is a stable and heritable epigenetic mark, and it plays an important role in regulation of gene expression and transposon silencing. Here we developed a CRISPR/dCpf1-based targeted demethylation system using the catalytic domain of the human demethylase TEN-ELEVEN TRANSLOCATION1 (TET1cd) and a SunTag system. The SunTag-dCpf1-TET1cd system is able to achieve targeted DNA demethylation and ur-regulate gene expression when guided to the FWA or CACTA1 loci in Arabidopsis thaliana. Our study provides tools for targeted removal of DNA cytosine methylation, and activation of protein-coding genes or transposons expression.

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"TRADES": Targeted RNA demethylation by Suntag system

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TRADES: Targeted RNA Demethylation by SunTag System.

Project description:N6-methyladenosine (m6A) is rapidly being studied and uncovered to play a significant role in various biological processes as well as in RNA fate and functions, while the effects of defined m6A sites are rarely characterized for the lack of convenient tools. To provide an applicable method to remove m6A modification at specific loci, an m6A editing system called "targeted RNA demethylation by SunTag system (TRADES)" is engineered. In this system, the targeting element dCas13b is fused to ten copies of GCN4 peptides which can recruit multiple scFv-fusion RNA demethylase to demethylate the target m6A site. Owing to this design, TRADES is more flexible to the indistinct m6A sites for its wide editing window. By site-specific demethylation of messenger RNA (mRNA) SON A2699, the lifetime of SON RNA is successfully prolonged in HeLa cells. Meanwhile, TRADES negligibly influences the lifetime of other non-targeted transcripts. TRADES also can regulate the gene expression of target transcript in an m6A-dependent manner. Moreover, the interference occuring for HBV and HIV replications demonstrates that the TRADES system holds potential in viral life cycle regulation and clinical applications.

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