Project description:Understanding the functional role of RNA modifications and to what extent they might regulate mature miRNA function and their secretion in extracellular vesicles (EVs) is unknown. Analysis of RNA content across a variety of EVs has shown differential miRNA enrichment when compared to parental cell expression patterns. Whether this is due to differential stability, specific regulation of miRNA export, or other mechanisms is unclear. Here, we tested whether RNA base modifications and recognition of those modifications might underlie differential miRNA export into EVs derived from KRAS mutant cell lines. We found that decreased levels of Mettl3, a writer of N6-methyladenosine (m6A) modification, altered extracellular transfer of miRNAs containing consensus sequences for m6A. Further, EVs prepared from cells expressing shRNAs against Mettl3 were incapable of conferring colony growth in 3D to wild type KRAS cells. Our data indicate that m6A modification plays an important role in miRNA export to EVs.
Project description:To evaluate the effects of ALKBH5 KD on the mRNA stability, we conducted RNA-seq of mRNA samples enriched from AML cells with ALKBH5 knockdown and actinomycin D treatment for different time
Project description:To identify the potential mRNA targets of ALKBH5, we conducted RNA-seq of mRNA samples enriched from AML cells with ALKBH5 knockdown
Project description:Understanding the functional role of RNA modifications and to what extent they might regulate mature miRNA function and their secretion in extracellular vesicles (EVs) is unknown. Analysis of RNA content across a variety of EVs has shown differential miRNA enrichment when compared to parental cell expression patterns. Whether this is due to differential stability, specific regulation of miRNA export, or other mechanisms is unclear. Here, we tested whether RNA base modifications and recognition of those modifications might underlie differential miRNA export into EVs derived from KRAS mutant cell lines. We found that decreased levels of Mettl3, a writer of N6-methyladenosine (m6A) modification, altered extracellular transfer of miRNAs containing consensus sequences for m6A. Further, EVs prepared from cells expressing shRNAs against Mettl3 were incapable of conferring colony growth in 3D to wild type KRAS cells. Our data indicate that m6A modification plays an important role in miRNA export to EVs.
Project description:To understand the effects of VRK1 knockdown on global phospho- and total- proteomics we profiled the U251MG VRK2-low and VRK2-high cell lines at 5- and 7-days post-doxycycline treatment.
Project description:we find METTL3 associates with polyribosomes and promotes translation. METTL3 depletion inhibits translation, and both wild-type and catalytically inactive METTL3 promote translation when tethered to the 3' untranslated region (UTR) of a reporter mRNA. Mechanistically, METTL3 enhances mRNA translation through an interaction with the translation initiation machinery. m6A seq in A549 and H1299 cells, RNA seq in METTL3 knockdown cells
Project description:To study the effects of hypoxia-related m6A modification which were mediated by ALKBH5 in pancreatic cancer, we established the ALKBH5 knockdown cell lines in PANC-1 cells. And then, the ALKBH5 knockdown cell lines were cultured under hypoxia for 48 h.
Project description:To identify the potential mRNA targets of ALKBH5, we conducted m6A-seq with mRNA samples enriched from AML cells with ALKBH5 knockdown or overexpression