1. DNA methylation is a critical epigenetic modification that silences gene transcription, participates in X-chromosome inactivation in females, and regulates genomic imprinting.
2. We have devised a method to inhibit transcriptional initiation by constructing short methylated oligonucleotides which induce DNA methylation at specific loci.
3. The methodology by which we devise these oligonucleotides is described, using oligonucleotides directed against the oncogene, Bcl-2.
4. The human Bcl-2 gene contains two promoters, each of which contains a CpG island in its core region. Oligonucleotides are designed which can inhibit Bcl-2 transcription and lead to decreased mRNA and protein in vitro. When compared to standard anti-sense oligonucleotide action, these methylated oligonucleotides are far more sensitive and potentially, longer acting.
5. In principle, using this methodology, it should be possible to design methylated oligonucleotides that can methylate CpG islands and thereby downregulate any gene.
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ACKNOWLEDGMENTS
We thank Dr Xiaoming Yao and Dr Jie Wu for assistance with the laboratory work in this project. Supported by NIH grant DK36054 and the Research Service of the Department of Veterans Affairs to ARH, and Department of Defense Grant no. W81XWH-04-1-0597 and NIH Grant no. R43 CA86664-01 to JFH.
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Hoffman, A.R., Hu, J.F. Directing DNA Methylation to Inhibit Gene Expression. Cell Mol Neurobiol 26, 423–436 (2006). https://doi.org/10.1007/s10571-006-9057-5
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DOI: https://doi.org/10.1007/s10571-006-9057-5