High levels of protein expression using different mammalian CMV promoters in several cell lines

CNS-directed gene therapy for the treatment of neurologic and somatic mucopolysaccharidosis type II (Hunter syndrome)

“…Later, the same group tested the i.v. administration of AAV5 vectors to newborn (P2) MPSII mice; IDS expression from this vector was driven by the strong viral promoter CMV (65). Although in this case, the levels of circulating and brain IDS were much lower than those observed in their previous work, reaching only 1% to 2% of WT IDS activity in the brain, and the impact on neurological disease seemed to be greater (60), likely due to the treatment of animals at an age at which the disease had barely manifested.…”

Effect of promoter strength on protein expression and immunogenicity of an HSV-1 amplicon vector encoding HIV-1 Gag

“…Our experiments were focused principally on the comparative analysis of different promoter elements in the context of the HSV-1 amplicon vector system, since the optimization of transcriptional control sequences has been shown to substantially improve the immunogenicity of both DNA plasmid and adenovirally-vectored vaccines [20,23,32,35,36,[42][43][44][45]. In HIV-1 DNA vaccines, the use of the hybrid CMV-I transcriptional control element resulted in improved immunogenicity, compared to vectors that contained a simple CMV promoter without Intron A [23,42,43].…”

“…Promoters chosen for this study included the ubiquitously active human cytomegalovirus immediate early (CMV) promoter, as well as three hybrid regulatory elements derived from this promoter. These hybrid promoters contained combinations of the CMV promoter/enhancer together with (i) its associated Intron A element (CMV-I) [21,22,[32][33][34], (ii) the regulatory R region from the long terminal repeat (LTR) of the human T-cell leukemia virus type 1 (CMV-R) [20], and (iii) the chicken β-actin promoter and the woodchuck hepatitis virus regulatory element (CAG) [19]. Each of these composite CMV-based promoters has been reported to mediate improved levels of gene expression and/or enhanced immune responses to encoded antigens, when compared to the basic CMV promoter [35,36].…”

Utilization of non‐AUG initiation codons in a flow cytometric method for efficient selection of recombinant cell lines

“…To address the need for greater yields, several different strategies have been pursued. These include utilizing stronger promoters in expression vectors (Gaillet et al, 2010;Prentice et al, 2007;Running Deer and Allison, 2004;Xia et al, 2006), better post-transfection selection systems (Sautter and Enenkel, 2005;van Blokland et al, 2007), and more efficient strategies for selecting clones with high expression levels (Bailey et al, 2002;Bohm et al, 2005;Brezinsky et al, 2003;Browne and Al-Rubeai, 2007;Caron et al, 2009). With respect to clone selection strategies, we and others have shown that IRES mediated expression of a reporter in conjunction with flow cytometry or magnetic separation methods can effectively isolate high expressing clones without requiring an antibody specific for the recombinant protein of interest (DeMaria et al, 2007;Gaines and Wojchowski, 1999;Liu et al, 2000;Sleiman et al, 2008).…”