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Dinh et al. Silence 2013, four:1 http://www.silencejournal/content/4/1/METHODSOpen AccessGeneration of a luciferase-based reporter for CHH and CG DNA methylation in Arabidopsis thalianaThanh Theresa Dinh1,two, Michael O’Leary1,4, So Youn Won1, Shengben Li1, Lorena Arroyo1, Xigang Liu1, Andrew Defries1,2, Binglian Zheng1,5, Sean R Cutler1 and Xuemei Chen1,3*AbstractBackground: DNA methylation guarantees genome integrity and regulates gene expression in diverse eukaryotes. In Arabidopsis, methylation occurs in 3 sequence contexts: CG, CHG and CHH. The initial establishment of DNA methylation at all three sequence contexts happens through a method generally known as RNA-directed DNA methylation (RdDM), in which compact RNAs bound by Argonaute4 (AGO4) guide DNA methylation at homologous loci via the de novo methyltransferase DRM2. After established, DNA methylation at each of the three sequence contexts is maintained by way of diverse mechanisms.Vardenafil hydrochloride Though some players involved in RdDM and maintenance methylation have been identified, the underlying molecular mechanisms usually are not completely understood.Thyrotropin To aid the complete identification of players in DNA methylation, we generated a transgenic reporter method that permits genetic and chemical genetic screens in Arabidopsis.PMID:23008002 Outcomes: A dual 35S promoter (d35S) driven luciferase (LUC) reporter was introduced into Arabidopsis and LUCL, a line using a low basal degree of luciferase activity, was obtained. LUCL was discovered to be a multi-copy, single-insertion transgene that includes methylated cytosines in CG, CHG and CHH contexts, using the highest methylation in the CG context. Methylation was present throughout the promoter and LUC coding region. Therapy with an inhibitor of cytosine methyl.