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Nucleic Acids Research 2006 34(13):e96; doi:10.1093/nar/gkl527
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Published online 7 August 2006

Nucleic Acids Research, 2006, Vol. 34, No. 13 e96
© 2006 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commerical use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

Engineering a high-affinity methyl-CpG-binding protein

Helle F. Jørgensen*, Karen Adie, Pascal Chaubert1 and Adrian P. Bird*

The Wellcome Trust Centre for Cell Biology, University of Edinburgh King's Buildings, Mayfield Road, Edinburgh EH9 3JR, UK 1 Institut Universitaire de Pathologie, Centre Hospitalier Universitaire Vaudois Bugnon 25, Lausanne CH-1011, Switzerland

*To whom correspondence should be addressed at The Wellcome Trust Centre for Cell Biology, University of Edinburgh, Michael Swann Building, Mayfield Road, Edinburgh EH9 3JR, UK. Tel: +44 131 650 8695; Fax: +44 131 650 5379; Email: A.bird{at}ed.ac.uk

Received May 3, 2006. Revised July 6, 2006. Accepted July 9, 2006.

Core members of the MBD protein family (MeCP2, MBD1, MBD2 and MBD4) share a methyl-CpG-binding domain that has a specific affinity for methylated CpG sites in double-stranded DNA. By multimerizing the MDB domain of Mbd1, we engineered a poly-MBD protein that displays methyl-CpG-specific binding in vitro with a dissociation constant that is >50-fold higher than that of a monomeric MBD. Poly-MBD proteins also localize to methylated foci in cells and can deliver a functional domain to reporter constructs in vivo. We propose that poly-MBD proteins are sensitive reagents for the detection of DNA methylation levels in isolated native DNA and for cytological detection of chromosomal CpG methylation.

*Correspondence may also be addressed to Helle F. Jørgensen. Tel: +44 208 383 8286; Fax: +44 208 383 8338; Email: helle.jorgensen{at}csc.mrc.ac.uk

Present address: Helle F. Jørgensen, Lymphocyte Development Group, MRC Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Hospital, Du Cane Road, London W12 0NN, UK


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