5-methylcytosine Modification

5-methylcytosine modification is a chemical alteration in which a methyl group is added to the fifth carbon of cytosine, a nucleic acid base, and it plays important roles in regulating genetic information. In DNA, DNA methyltransferases transfer methyl groups from S-adenosylmethionine to cytosine, often at cytosine-guanine sites, influencing chromatin structure and gene transcription without changing the underlying DNA sequence. This epigenetic mark contributes to development, genomic stability, and cell-specific gene expression. Abnormal methylation patterns are associated with cancer, imprinting disorders, and other diseases, making 5-methylcytosine analysis valuable in developmental biology, disease research, and biomarker development.

5-methylcytosine Modification - Related Videos

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JoVE EoE - Blot Techniques

5-Methylcytosine Dot Blot to Determine the Extent of DNA Methylation

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2025

This video describes the technique of dot-blot to determine the level of DNA methylation in human chondrocytes in vitro via the blotting of DNA samples on a nylon membrane and the subsequent visualization using monoclonal antibodies. This helps to determine the chondrocytes' phenotype at various stages in culture.

Research

JoVE Journal - Developmental Biology
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Immunohistochemical Detection of 5-Methylcytosine and 5-Hydroxymethylcytosine in Developing and Postmitotic Mouse Retina

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Cited by 16 •

2018

The objective of this report is to describe the protocol for robust immunohistochemical detection of epigenetic markers, 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) in developing and postmitotic mouse retina.

Efficient Purification and LC-MS/MS-based Assay Development for Ten-Eleven Translocation-2 5-Methylcytosine Dioxygenase

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Cited by 5 •

2018

Here, we present a protocol for an efficient single step purification of the active untagged human ten-eleven translocation-2 (TET2) 5-methylcytosine dioxygenase using ion-exchange chromatography and its assay using a liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based approach.

Research

JoVE Journal - Genetics
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Methylated RNA Immunoprecipitation Assay to Study m5C Modification in Arabidopsis

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Cited by 18 •

2020

The methylated RNA immunoprecipitation assay is an antibody-based method used to enrich for methylated RNA fragments. Coupled with deep sequencing, it leads to the identification of transcripts carrying the m5C modification.

A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro

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Cited by 17 •

2017

We present a method to quantify DNA methylation based on the 5-methylcytosine (5-mC) dot blot. We determined the 5-mC levels during chondrocyte dedifferentiation. This simple technique could be used to quickly determine the chondrocyte phenotype in ACI treatment.

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