Role and Regulation of DNA Methylation in Cellular Reprogramming
Biochemistry
Final Report Abstract
We could show that global DNA hypomethylation prevents consolidation of differentiation programs and allows reversion to the embryonic stem cell state. We developed a novel recombinant transcription (designer TALE) to specifically activate the oct4 pluripotency gene and demonstrated targeted transcriptional activation of silent oct4 pluripotency gene by combining designer TALEs and inhibition of epigenetic modifiers. We developed two new methods to quantify and map this new DNA base (5 hydroxymethylcytosine, 5hmC) in pluripotency and differentiation. We could show that the new DNA base 5hmC is recognized by the multi-domain protein Uhrf1, which is essential for the maintenance of DNA methylation in stem cells. In summary, we developed novel tools to study DNA modifications that are now widely used in the scientific community and contributed to a better understanding of the role of DNA modifications during development, differentiation and reprogramming.
Publications
- (2010). Differentiation and large scale spatial organization of the genome. Curr Opin Genet Dev. 20, 562-569
Joffe, B., Leonhardt, H. and Solovei, I.
(See online at https://doi.org/10.1016/j.gde.2010.05.009) - (2010). Sensitive enzymatic quantification of 5-hydroxymethylcytosine in genomic DNA. Nucl. Acids Res. 38, e181
Szwagierczak, A., Bultmann, S., Schmidt, C.S., Spada, F. and Leonhardt, H.
(See online at https://doi.org/10.1093/nar/gkq684) - (2011). Characterization of PvuRts1I endonuclease as a tool to investigate genomic 5 hydroxymethylcytosine. Nucl. Acids Res. 39, 5149-56
Szwagierczak, A., Brachmann, A., Schmidt, C.S., Bultmann, S., Leonhardt, H. and Spada, F.
(See online at https://doi.org/10.1093/nar/gkr118) - (2011). Generation and characterization of rat and mouse monoclonal antibodies specific for MeCP2 and their use in X-inactivation studies. PLoS ONE 6: e26499
Jost, K. L., Rottach, A., Milden, M., Bertulat, B., Becker, A., Wolf, P., Sandoval, J., Petazzi, P., Huertas, D., Esteller, M., Kremmer, E., Leonhardt, H. and Cardoso, M. C.
(See online at https://doi.org/10.1371/journal.pone.0026499) - (2011). Recognition of 5-hydroxymethylcytosine by the Uhrf1 SRA domain. PLoS ONE 6: e21306
Frauer, C., Hoffmann, T. Bultmann, S., Casa, V., Cardoso, M. C., Antes, I. and Leonhardt, H.
(See online at https://doi.org/10.1371/journal.pone.0021306) - (2012). Global DNA Hypomethylation Prevents Consolidation of Differentiation Programs and Allows Reversion to the Embryonic Stem Cell State. PLoS One 2012 Dez, 7, e52629
Schmidt, C.S., Bultmann, S., Meilinger, D., Zacher, B., Tresch, A., Maier, K.C., Peter, C., Martin, D.E., Leonhardt, H. and Spada, F.
(See online at https://doi.org/10.1371/journal.pone.0052629) - (2012). Targeted transcriptional activation of silent oct4 pluripotency gene by combining designer TALEs and inhibition of epigenetic modifiers. Nucleic Acids Res 40, 5368-77
Bultmann, S., Morbitzer, R., Schmidt, C.S., Thanisch, K., Spada, F., Elsaesser, J., Lahaye, T. and Leonhardt, H.
(See online at https://doi.org/10.1093/nar/gks199)
