Project description:Both benign nevi and melanomas exhibit high rates of activating mutations in BRAF kinase, suggesting that this event is important for melanocyte proliferation and melanoma initiation in vivo; however, the precise mechanism of BRAF kinase action in these lesions remains to be elucidated. We have used comprehensive gene expression profiling to evaluate the downstream effectors of activated BRAFV600E kinase in primary human melanocytes and identified a dominant proliferative target gene signature for BRAF kinase in these cells. We used microarrays to detail the global gene expression pattern change induced by the activation of RAS/BRAF/MEK/ERK signaling pathway and identified distinct classes of up-regulated genes during this process. Keywords: Genetic modification
Project description:Both benign nevi and melanomas exhibit high rates of activating mutations in BRAF kinase, suggesting that this event is important for melanocyte proliferation and melanoma initiation in vivo; however, the precise mechanism of BRAF kinase action in these lesions remains to be elucidated. We have used comprehensive gene expression profiling to evaluate the downstream effectors of activated BRAFV600E kinase in primary human melanocytes and identified a dominant proliferative target gene signature for BRAF kinase in these cells. We used microarrays to detail the global gene expression pattern change induced by the activation of RAS/BRAF/MEK/ERK signaling pathway and identified distinct classes of up-regulated genes during this process. Experiment Overall Design: Primary human melanocytes were isolated from neonatal foreskins. Melanocytes isolated from five individual donors were pooled together in order to reduce genetic variations and transduced by BRAFV600E expressing lentivirus and control virus with MOI 10 respectively. Cells were harvested 5 days following infection and total RNA was extracted and purified by RNeasy kit by following manufacturerâs protocol. RNA quality check, double strand complementry DNA synthesis, hybridization with Human Genome U133 Plus 2.0 Array Chips (Affymetrix Inc. Santa Clara, CA), initial data extraction, normalization using RMAExpress software.
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs.
Project description:Transcriptional profiling of human mesenchymal stem cells comparing normoxic MSCs cells with hypoxic MSCs cells. Hypoxia may inhibit senescence of MSCs during expansion. Goal was to determine the effects of hypoxia on global MSCs gene expression.