title: Improved antibiotic-free DNA vaccine vectors utilizing a novel RNA based plasmid selection system.
abstract: To ensure safety, regulatory agencies recommend elimination of antibiotic resistance markers from therapeutic and vaccine plasmid DNA vectors. Here, we describe the development and application of a novel antibiotic-free selection system. Vectors incorporate and express a 150 bp RNA-OUT antisense RNA. RNA-OUT represses expression of a chromosomally integrated constitutively expressed counter-selectable marker (sacB), allowing plasmid selection on sucrose. Sucrose selectable DNA vaccine vectors combine antibiotic-free selection with highly productive fermentation manufacturing (>1g/L plasmid DNA yields), while improving in vivo expression of encoded proteins and increasing immune responses to target antigens. These vectors are safer, more potent, alternatives for DNA therapy or vaccination.
authors: Nature Technology Corporation, 4701 Innovation Drive, Lincoln, NE 68521, USA.
authors: Jeremy
authors: J
authors: Luke
authors: Aaron E
authors: AE
authors: Carnes
authors: Clague P
authors: CP
authors: Hodgson
authors: James A
authors: JA
authors: Williams
mesh_terms: D000818:Animals; D001287:Attachment Sites, Microbiological; D002460:Cell Line; D005260:Female; D005285:Fermentation; D005822:Genetic Vectors / Q000096:biosynthesis*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012313:RNA / Q000235:genetics*; D014162:Transfection; D019444:Vaccines, DNA / Q000096:biosynthesis* / Q000276:immunology
chemical_list: D019444:Vaccines, DNA; D012313:RNA