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Open Access Methodology article

Proximity ligation in situ assay for monitoring the global DNA methylation in cells

Eric Hervouet12, Philippe Hulin23, François M Vallette12 and Pierre-François Cartron12*

Author Affiliations

1 Centre de Recherche en Cancérologie Nantes-Angers, INSERM, U892, Equipe Apoptose et Progression Tumorale, Equipe labellisée Ligue Nationale Contre le Cancer. 8 quai moncousu, BP7021, 44007 Nantes, France

2 Université de Nantes, Faculté de Médecine, Département de Recherche en Cancérologie, IFR26, F-4400, Nantes, France

3 PICell, Plateau technique d'imagerie cellulaire - IFR26 - Centre de Recherche en Cancérologie Nantes-Angers, INSERM, U892. F-4400, Nantes, France

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BMC Biotechnology 2011, 11:31  doi:10.1186/1472-6750-11-31

Published: 6 April 2011

Abstract

Background

DNA methylation has a central role in the epigenetic control of mammalian gene expression, and is required for X inactivation, genomics imprinting and silencing of retrotransposons and repetitive sequences. Thus, several technologies have been developed to measure the degree of DNA methylation.

Results

We here present the development of the detection of protein-protein interactions via the adaptation of the proximity ligation in situ technology to evaluate the DNA methylation status in cells since the quantification of Dnmt1/PCNA interaction in cells reflects the degree of DNA methylation.

Conclusion

This method being directly realizable on cells, it appears that it could suggest a wide range of applications in basic research and drug development. More particularly, this method is specially adapted for the investigations realized from a weak quantity of biologic materiel such as stem cells or primary cultured tumor cells for examples.

Keywords:
Global DNA methylation; method; Dnmt1/PCNA; proximity ligation in situ technology