RRC ID 73774
Author Otake K, Kugou K, Robertlee J, Ohzeki JI, Okazaki K, Hanano S, Takahashi S, Shibata D, Masumoto H.
Title De novo induction of a DNA-histone H3K9 methylation loop on synthetic human repetitive DNA in cultured tobacco cells.
Journal Plant J
Abstract Genetic modifications in plants are crucial tools for fundamental and applied research. Transgene expression usually varies among the independent lines or their progeny and is associated with the chromatin structure of the insertion site. Strategies based on understanding how to manipulate the epigenetic state of the inserted gene cassette would help ensure transgene expression. Here, we report a strategy for chromatin manipulation by artificial tethering of epigenetic effectors to a synthetic human centromeric repetitive DNA (alphoid DNA) platform in plant BY-2 culture cells. By tethering DNA-methyltransferase (Nicotiana tabacum DRM1), we effectively induced DNA methylation and histone methylation (H3K9me2) on the alphoid DNA platform. Tethering of the Arabidopsis SUVH9, which has been reported to lack histone methyltransferase activity, also induced a similar epigenetic state on the alphoid DNA in BY-2 cells, presumably by activating the RNA-dependent DNA methylation (RdDM) pathway. Our results emphasize the interplay between DNA and histone methylation mechanisms intrinsic to plant cells. We also found that once epigenetic modification states were induced by tethering of either DRM1 or SUVH9, the modification was maintained even when direct tethering of the effector was inhibited. Our system enables analysis of more diverse epigenetic effectors and can support elucidation of chromatin assembly mechanisms in plant cells.
Published 2023-2-24
DOI 10.1111/tpj.16164
PMID 36825961
IF 6.141
Arabidopsis / Cultured plant cells, genes rpc00001