ADAR1 overexpression in MDA-MB-231 cells

Abstract: RNA editing has emerged as novel mechanisms involved in cancer progression. Despite the phenotypic implications of ADAR in several cancer models, the role of ADAR on DNA damage response (DDR) and proliferation in breast cancer (BC) has not been fully addressed. To evaluate the effect of ADAR expression, MDA-MB-231 cells were transduced (MOI:200) with commercial pre-package adenoviral particles coding for ADARp110 or transduced (MOI:200) with eGFP control adenovirus. 16 h after transduction media was changed and RNA were extracted 48 h after transduction. ADARp110 overexpression produces a significant increase in transcripts related with cell cycle and proliferative pathways, in addition to other pathways related to IL10 signaling, among others. In other hand 657 sites significantly increase their editing in MDA-MB-231 OV cells at site level comparison. Conversely, transcripts involved in Generic Transcription, cell cycle, HIV life Cycle related pathways and Apoptosis suffer an increased editing ratio, suggesting that ADAR activity could be is in the regulation of the targets that bears significant changes in their editing pattern after ADAR manipulation Overall design: MDA-MB-231 cells were transduced (MOI: 200) with commercial pre-package adenoviral particles (VectorBuilder Inc, Shenandoah, TX, USA. VB170930-1035dgj) coding for ADARp110 (based on NM_001025107.2) or transduced (MOI: 200) with eGFP control adenovirus (VB150925-10024 vector). 16 h after transduction media was changed. Cells where extracted 48h after transduction using Qiagen RNeasy columns. RNA-seq libraries were sequenced using a Hiseq4000 (BGI, Korea). Fastq files were aligned using STAR and hg19, counting transcripts using QuickRNASeq. Differential expression analysis was performed using DEseq2 software following standard recommendations.