NTC8685-eRNA41H Nanoplasmid vector質粒載體

NTC8681, NTC8682, NTC8684 and NTC8685 vector family Introduction. NTC8681, NTC8682, NTC8684 and NTC8685 plasmids are antibiotic-free vectors optimized to combine maximal eukaryotic gene expression with superior bacterial manufacturing yields. These plasmids were specifically designed as safe minimalized antibiotic-free selection vectors for the expression of recombinant proteins in mammalian cells. This may be for protein production, gene therapy or induction of neutralizing immune responses by genetic immunization. The vectors combine minimal prokaryotic sequences including an antibiotic-free sucrose selectable marker. The vectors also contain a novel chimeric promoter that directs superior mammalian cell expression (Luke et al. 2009, 2011b). The vectors are available in four versions. NTC8681 targets encoded protein to the endosome using an optimized human lysosomal-associated membrane protein 1 (Lamp1) targeting tag. NTC8682 targets encoded protein into the secretory pathway using an optimized tissue plasminogen activator (TPA) signal peptide. NTC8684 targets proteins to the proteosome by fusion C-terminal to a destabilizing UbiquitinA76 tag. NTC8685 expresses encoded protein native without targeting sequences. The vectors were designed to be responsive to Food and Drug Administration (FDA) regulatory guidance s regarding DNA Vaccine vector composition (FDA 1996, FDA 2007; reviewed in Williams et al. 2009a). All sequences that were not essential for Escherichia coli plasmid replication or mammalian cell expression of the target gene were eliminated. Synthetic eukaryotic mrna leader and terminators were utilized in the vector design to limit DNA sequence homology with the human genome to reduce the possibility of chromosomal integration. The vectors encode a consensus Kozak translation initiation sequence and ATG start codon. Target gene expression is driven from an optimized chimeric promoter-intron (SV40- CMV-HTLV-1 R synthetic intron). The boundary between the CMV promoter and the SV40 enhancer has been optimized resulting in dramatically improved expression in mammalian cells (Luke et al. 2011b). This chimeric CMV promoter achieves significantly higher expression levels than traditional human cytomegalovirus (CMV) promoter based vectors (Luke et al. 2009, 2011b). NTC8681, NTC8682, NTC8684 and NTC8685 vectors also incorporate the adenoviral serotype 5 VA RNAI (VA1) transient expression enhancer. VA1 further improves eukaryotic expression without affecting Escherichia coli production yields (Carnes et al. 2010, 2011). NTC86-series erna41h Targeting RIG-I Activating Expression Vectors 4

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