Project description:Purpose: We examined how transcriptional state changes relate to clonal selection as BRAF inhibitor resistance develops in BRAF-mutated myeloma. Methods: To this end we generated three single-cell clones from U266, a BRAFK601N -mutated myeloma cell line and DP6- a BRAFV600E myeloma cell line. All three U266 and DP6 clones were subjected to long-term dabrafenib treatment at their established IC50 doses (U266: 10uM, DP6 1nM). Bulk RNA-seq was performed before treatment, day 7, day 14, day 42 and at time of resistance. Results: Transcriptional adaptation after seven days was homogeneous for all clones, but was different across both cell lines. Oxidative phosphorylation (OxPhos) emerged as the most consistently enriched signaling pathway in persistent cells from both cell lines as compared to baseline. Conclusions: BRAF inhibition in BRAF-mutated myeloma cells leads to transcriptional reprogramming with induction of OxPhos-related genes within a brief period of time.
Project description:We performed MeRIP-seq on (1) two human epidermis tissue and HaCAT (human keratinocyte cell line) cells; (2) HaCAT cell lines with or without ALKBH5 knockdown by siRNA in duplicate experiments.
Project description:Purpose and Methods: To profile m6A modifications on both the viral genome RNA and cellular transcripts, we used Methylated RNA immunoprecipitation sequencing (MeRIP-seq) on ZIKV SZ1901-infected human hepatocarcinoma cell line Huh7. Results and conclusions: Using MeRIP-seq, we identified a unique m6A map in the genome of ZIKV strain SZ1901 that is different from all previous isolates. Our study describes unique viral and host m6A profiles in contemporary ZIKV-infected Huh7 cells, highlighting the complexity and importance of m6A modification during viral infection.
Project description:To determine the targets underlying ALKBH5 during head and neck squamouse cell carcinoma progression, Methylated RNA immunoprecipitation (MeRIP) with an m6A specific antibody followed by RNA sequencing (MeRIP-seq) and next generation sequencing were combined to screen the potential targets haboring m6A modificatios and mRNA level alteration after ALKBH5 knockdown in a HNSCC cell line.
Project description:To determine the targets underlying ALKBH5 during head and neck squamouse cell carcinoma progression, Methylated RNA immunoprecipitation (MeRIP) with an m6A specific antibody followed by RNA sequencing (MeRIP-seq) and next generation sequencing were combined to screen the potential targets haboring m6A modificatios and mRNA level alteration after ALKBH5 knockdown in a HNSCC cell line.
Project description:Melphalan-induced modulation of miR-221/222 levels in MM cells. Melphalan-resistant U266/LR7 cells showed the highest induction of miR-221/222 after drug exposure. To study the transcriptome perturbation induced in MM cells following the combination of miR-221/222 inhibitors plus melphalan we used the whole gene expression data total RNA was obtained after single or combination treatment of the Melphalan-resistant U266/LR7 cells and the parental cell line U266/s
Project description:Significance of RNA methylation in the context of HIV-1 infection in human T cells MeRIP-Seq of Control and HIV-infected MT4 T-Cells