title: Evaluating regulatory elements of human cytomegalovirus major immediate early gene for enhancing transgene expression levels in CHO K1 and HEK293 cells.
abstract: The upstream regulatory sequence (URS), NF1 region, enhancer, promoter, 1st exon, and intron A of human cytomegalovirus major immediate early gene (hCMV MIE) are evaluated for enhancing transient and stable gene expression levels in two industrial cell lines, CHO K1 and HEK293 using firefly luciferase (Fluc) and erythropoietin (EPO). As compared to the control vector which only contains the enhancer and promoter (EP), vectors containing the 1st exon (EPE) and intron A (EPEI) enhance transient expression levels of the two proteins by approximately 2.5- to 4.3-fold in the two cell lines. Addition of NF1 and URS to EP (NEP and UNEP) or EPEI (NEPEI and UNEPEI) results in a lesser effect on the expression. In stable transfections, UNEPEI provides the highest expression level in CHO K1 cells, yielding approximately 4.0-fold increase in Fluc expression and 2.5-fold increase in EPO expression. In HEK293 cells, EPE is the best and enhances Fluc and EPO expression by more than 2.0-fold. Such information is valuable for the development of optimal vectors to enhance transient and stable production of recombinant proteins in CHO K1 and HEK293 cells.
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore.
authors.lastname: Mariati
authors.forename: Yi Kai
authors.initials: YK
authors.lastname: Ng
authors.forename: Sheng-Hao
authors.initials: SH
authors.lastname: Chao
authors.forename: Miranda G S
authors.initials: MG
authors.lastname: Yap
authors.forename: Yuansheng
authors.initials: Y
authors.lastname: Yang
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003587:Cytomegalovirus / Q000235:genetics*; D015870:Gene Expression; D017781:Genes, Immediate-Early / Q000235:genetics*; D006801:Humans; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D019076:Transgenes / Q000235:genetics*