Miller JL
National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health,
10 Center Drive, Building 10, Room 9N311, Bethesda, Maryland 20892
Tanzania
Short Communication
Puromycin Selection Confounds the RNA-Seq Profiles of Primary Human Erythroblasts
Author(s): Guo RL, Lee YT, Byrnes C and Miller JLGuo RL, Lee YT, Byrnes C and Miller JL
Lentiviral transduction followed by puromycin selection is a well-recognized procedure for gene transfer and expression experiments using a variety of cell types including human hematopoietic stem and progenitor cells. Despite its widespread application, research regarding the potential effects of bacterial puromycin N-acetyltransferase (pac) gene expression in mammalian cell cultures is incomplete. Here the potential for puromycin selection to affect transcriptome profiles was examined using a well-studied model for human erythropoiesis. Experiments were performed using primary CD34(+) cells from six adult healthy human donors transduced with two commercially available pac-encoding lentiviral vectors and compared to non-transduced control cells. RNA-Seq gene expression profiles were generated at the proerythroblast stage of differentiation, then differential gene expression was analy.. View More»
