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Single-cell m6A mapping in vivo using picoMeRIP-seq.
Yanjiao Li1,2, Yunhao Wang3,4, Maria Vera-Rodriguez1,5
1Department of Microbiology, Oslo University Hospital, Rikshospitalet, Oslo, Norway.
Nature Biotechnology
|June 22, 2023
Summary
We developed picogram-scale m6A RNA immunoprecipitation and sequencing (picoMeRIP-seq) to map RNA methylation in single cells and scarce samples. This method requires minimal RNA, overcoming limitations of current techniques for in vivo studies.
Area of Science:
- Molecular Biology
- Genomics
- Epigenetics
Background:
- Current N6-methyladenosine (m6A) mapping techniques require substantial RNA quantities.
- Existing methods are often restricted to cultured cell lines, limiting in vivo applications.
- There is a need for sensitive m6A detection in scarce biological samples.
Purpose of the Study:
- To develop a highly sensitive method for m6A mapping.
- To enable m6A profiling in single cells and low-input biological samples.
- To facilitate in vivo studies of m6A modification.
Main Methods:
- Optimized sample recovery and signal-to-noise ratio for immunoprecipitation.
- Developed picogram-scale m6A RNA immunoprecipitation and sequencing (picoMeRIP-seq).
- Utilized standard laboratory equipment for accessibility.
Main Results:
- Successfully mapped m6A at picogram RNA levels.
- Demonstrated the method's efficacy in titrations of poly(A) RNA and embryonic stem cells.
- Applied picoMeRIP-seq to single zebrafish zygotes, mouse oocytes, and embryos.
Conclusions:
- picoMeRIP-seq significantly reduces the RNA input requirement for m6A mapping.
- The method allows for in vivo m6A profiling in single cells and rare cell populations.
- picoMeRIP-seq provides a valuable tool for studying m6A dynamics in diverse biological contexts.
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