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A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues

A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues

Published on: December 5, 2016

N6-methyladenosine modulates long non-coding RNA in the developing mouse heart.

Siman Shen^1,2, Keyu Liu^1,2, Simeng Li^1,2

¹Department of Anesthesiology, Affiliated Hospital of Guangdong Medical University, Zhanjiang, China.

Cell Death Discovery

|July 20, 2022

Summary

N6-methyladenosine (m6A) modification of long non-coding RNAs (lncRNAs) is crucial for mouse cardiac development and myocardial regeneration. This study maps m6A modifications in lncRNAs during early cardiac development, revealing key regulatory roles.

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A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues

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Tissue-specific miRNA Expression Profiling in Mouse Heart Sections Using In Situ Hybridization

Tissue-specific miRNA Expression Profiling in Mouse Heart Sections Using In Situ Hybridization

Published on: September 15, 2018

Area of Science:

Molecular Biology
Cardiovascular Research
Epigenetics

Background:

Long non-coding RNAs (lncRNAs) are implicated in myocardial regeneration.
N6-methyladenosine (m6A) modification influences lncRNA stability and biological processes.
The m6A landscape of lncRNAs in mouse cardiac development is largely unknown.

Purpose of the Study:

To investigate stage-specific m6A modifications in cardiac lncRNAs during mouse development.
To reveal the role of m6A-modified lncRNAs in cardiac development and regeneration.
To identify novel lncRNAs involved in cardiac regeneration through m6A modification.

Main Methods:

Performed Methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA sequencing (RNA-seq) on mouse heart tissues at postnatal days P1, P7, and P28.
Utilized METTL3 silencing in neonatal cardiomyocytes in vitro to assess the functional role of m6A in regulating lncRNA levels.
Employed MeRIP-quantitative PCR (qPCR) to validate methylation status of specific lncRNAs.

Main Results:

Significant differences in the distribution and abundance of lncRNA m6A modifications were observed between P1 and P7 mouse hearts.
lncGm15328 and lncRNA Zfp597 showed the most differential methylation at P1-P7 and were not previously linked to cardiac regeneration.
Differential m6A modifications were enriched in pathways related to cardiomyocyte proliferation, suggesting a role in cardiac development and regeneration.

Conclusions:

m6A modification of lncRNAs plays a significant role in mouse cardiac development.
Specific m6A-modified lncRNAs, such as lncGm15328 and lncRNA Zfp597, may act as key regulators in myocardial regeneration.
These findings provide novel insights into the epigenetic regulation of cardiac development and the potential for therapeutic interventions in myocardial regeneration.