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GLORI for absolute quantification of transcriptome-wide m6A at single-base resolution

ABSTRACT: GLORI for absolute quantification of transcriptome-wide m6A at single-base resolution

PROVIDER: PRJNA978429 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24791673.fastq.gz	Fastqsanger.gz
	SRR24791674.fastq.gz	Fastqsanger.gz

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Mild and ultrafast GLORI enables absolute quantification of m6A methylome from low-input samples

Project description:Methods that enable absolute quantification of N6-methyladenosine (m6A) RNA modification have emerged as powerful tools in the field of epitranscriptomics. We previously reported GLORI, a chemical-assisted approach firstly achieved quantitatively transcriptome-wide m6A measurement at single-base resolution. Despite its advantages, GLORI suffers from lengthy reaction time and severe RNA degradation. Here, we present two updated GLORI approaches: GLORI 2.0 is an ultra-fast and mild version that preserves RNA integrity and enhances sensitivity for both transcriptome-wide and locus-specific m6A detection; GLORI 3.0 further utilizes a novel reverse transcription-silent carrier RNA to achieve high-quality m6A quantification from ~ 1,000 cells. Using limited RNA input extracted from single mouse dorsal hippocampus, we measure m6A methylome in the synaptic and cytoplasmic fractions and reveal a high modification level in synapse-related gene sets. We envision that the updated GLORI methods will greatly expand the applicability of absolute quantification of m6A in biology.

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Project description:We developed a quantitative method called GLORI to investigate m6A methylation in the mammalian transcriptome at single-base resolution.

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