Byrnes C
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»
