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Methylated RNA Immunoprecipitation Assay to Study m5C Modification in Arabidopsis.

Authors: Saplaoura, Eleftheria; Perrera, Valentina; Colot, Vincent; Kragler, Friedrich;

Methylated RNA Immunoprecipitation Assay to Study m5C Modification in Arabidopsis.

Abstract

Secondary base modifications on RNA, such as m5C, affect the structure and function of the modified RNA molecules. Methylated RNA Immunoprecipitation and sequencing (MeRIP-seq) is a method that aims to enrich for methylated RNA and ultimately identify modified transcripts. Briefly, sonicated RNA is incubated with an antibody for 5-methylated cytosines and precipitated with the assistance of protein G beads. The enriched fragments are then sequenced and the potential methylation sites are mapped based on the distribution of the reads and peak detection. MeRIP can be applied to any organism, as it does not require any prior sequence or modifying enzyme knowledge. In addition, besides fragmentation, RNA is not subjected to any other chemical or temperature treatment. However, MeRIP-seq does not provide single-nucleotide prediction of the methylation site as other methods do, although the methylated area can be narrowed down to a few nucleotides. The use of different modification-specific antibodies allows MeRIP to be adjusted for the different base modifications present on RNA, expanding the possible applications of this method.

Country
Italy
Keywords

5, Immunoprecipitation (IP), Base Sequence, Transcription, Genetic, Sequence Analysis, RNA, Arabidopsis, RNA sequencing, C), Methylation, Epitranscriptomic, Genetic, 5-methylcytosine (m, RNA, Plant, Issue 159, 5-methylcytosine (m; 5; C); Arabidopsis; Epitranscriptomics; Genetics; Immunoprecipitation (IP); Issue 159; MRNA; RNA methylation; RNA sequencing, MRNA, RNA methylation, 5-Methylcytosine, Immunoprecipitation, RNA, Messenger, Arabidopsi

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
Average
Average
Average
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