RT Journal Article
T1 DNA Methylation Editing by CRISPR-guided Excision of 5-Methylcytosine.
A1 Devesa-Guerra, Ivan
A1 Morales-Ruiz, Teresa
A1 Perez-Roldan, Juan
A1 Parrilla-Doblas, Jara Teresa
A1 Dorado-Leon, Macarena
A1 Garcia-Ortiz, Maria Victoria
A1 Ariza, Rafael R
A1 Roldan-Arjona, Teresa
K1 DNA demethylation
K1 DNA glycosylases
K1 TET dioxygenases
K1 Epigenetics
AB Tools for actively targeted DNA demethylation are required to increase our knowledge about regulation and specific functions of this important epigenetic modification. DNA demethylation in mammals involves TET-mediated oxidation of 5-methylcytosine (5-meC), which may promote its replication-dependent dilution and/or active removal through base excision repair (BER). However, it is still unclear whether oxidized derivatives of 5-meC are simply DNA demethylation intermediates or rather epigenetic marks on their own. Unlike animals, plants have evolved enzymes that directly excise 5-meC without previous modification. In this work, we have fused the catalytic domain of Arabidopsis ROS1 5-meC DNA glycosylase to a CRISPR-associated null-nuclease (dCas9) and analyzed its capacity for targeted reactivation of methylation-silenced genes, in comparison to other dCas9-effectors. We found that dCas9-ROS1, but not dCas9-TET1, is able to reactivate methylation-silenced genes and induce partial demethylation in a replication-independent manner. We also found that reactivation induced by dCas9-ROS1, as well as that achieved by two different CRISPR-based chromatin effectors (dCas9-VP160 and dCas9-p300), generally decreases with methylation density. Our results suggest that plant 5-meC DNA glycosylases are a valuable addition to the CRISPR-based toolbox for epigenetic editing.
PB Academic Press
YR 2020
FD 2020-02-07
LK http://hdl.handle.net/10668/15148
UL http://hdl.handle.net/10668/15148
LA en
NO Devesa-Guerra I, Morales-Ruiz T, Pérez-Roldán J, Parrilla-Doblas JT, Dorado-León M, García-Ortiz MV, et al. DNA Methylation Editing by CRISPR-guided Excision of 5-Methylcytosine. J Mol Biol. 2020 Mar 27;432(7):2204-2216
DS RISalud
RD Sep 27, 2025