DataSheet_1_Argonaute 4 as an Effector Protein in RNA-Directed DNA Methylation in Human Cells.pdf Kanwalat Chalertpet Piyapat Pin-on Chatchawit Aporntewan Maturada Patchsung Praewphan Ingrungruanglert Nipan Israsena Apiwat Mutirangura 10.3389/fgene.2019.00645.s001 https://frontiersin.figshare.com/articles/dataset/DataSheet_1_Argonaute_4_as_an_Effector_Protein_in_RNA-Directed_DNA_Methylation_in_Human_Cells_pdf/8667935 <p>DNA methylation of specific genome locations contributes to the distinct functions of multicellular organisms. DNA methylation can be governed by RNA-dependent DNA methylation (RdDM). RdDM is carried out by endogenous small-RNA-guided epigenomic editing complexes that add a methyl group to a precise DNA location. In plants, the Argonaute 4 (AGO4) protein is one of the main catalytic components involved in RdDM. Although small interfering RNA or short hairpin RNA has been shown to be able to guide DNA methylation in human cells, AGO protein-regulated RdDM in humans has not yet been evaluated. This study aimed to identify a key regulatory AGO protein involved in human RdDM by bioinformatics and to explore its function in RdDM by a combination of AGO4 knockdown, Alu small interfering RNA transfection, AGO4-expressing plasmid transfection, chromatin immunoprecipitation, cell-penetrating peptide-tagged AGO4 combined Alu single-guide RNA transfection, and methylation analyses. We found that first, human AGO4 showed stronger genome-wide association with DNA methylation than AGO1–AGO3. Second, endogenous AGO4 depletion demethylated DNA of known AGO4 bound loci. Finally, exogenous AGO4 de novo methylated the bound DNA sequences. Therefore, we discovered that AGO4 plays a role in human RdDM.</p> 2019-07-04 09:52:26 human Argonaute 4 human RNA-directed DNA methylation human RdDM CPP-AGO4 epigenomic editing