quantitative comparison of constitutive promoters in human es cells定量比较人类es细胞的组成型启动子.pdfVIP

Quantitative Comparison of Constitutive Promoters in Human ES cells 1 1 2 1 2 Karin Norrman , Yvonne Fischer , Blandine Bonnamy , Fredrik Wolfhagen Sand , Philippe Ravassard , Henrik Semb1* 1 Department of Laboratory Medicine, Lund Center for Stem Cell Biology and Cell Therapy, Lund University, Lund, Sweden, 2 Biotechnology and Biotherapy Laboratory, ˆ ´ ˆ ` Centre de Recherche de l’Institut du Cerveau et de la Moelle, CNRS UMR 7225, INSERM UMRS 975, University Pierre et Marie Curie, Hopital Pitie Salpetriere, Paris, France Abstract Background: Constitutive promoters that ensure sustained and high level gene expression are basic research tools that have a wide range of applications, including studies of human embryology and drug discovery in human embryonic stem cells (hESCs). Numerous cellular/viral promoters that ensure sustained gene expression in various cell types have been identified but systematic comparison of their activities in hESCs is still lacking. Methodology/Principal Findings: We have quantitatively compared promoter activities of five commonly used constitutive promoters, including the human b-actin promoter (ACTB), cytomegalovirus (CMV), elongation factor-1a, (EF1a), phosphoglycerate kinase (PGK) and ubiquitinC (UbC) in hESCs. Lentiviral gene transfer was used to ensure stable integration of promoter-eGFP constructs into the hESCs genome. Promoter activities were quantitatively compared in long term culture of undifferentiated hESCs and in their differentiated progenies. Co