title: Development of improved vectors for DNA-based immunization and other gene therapy applications.
abstract: Optimizing gene expression and delivery are necessary steps in the production of vectors for DNA-based immunization as well as for other gene therapy applications. A mouse muscle/reporter gene assay system was used to systematically improve a plasmid DNA vector. The optimized vector VR1255 contained: (1) CMV promoter and enhancer; (2) CMV IE Intron A; (3) kanamycin resistance gene; (4) deleted SV40 origin of replication; (5) optimized lux coding region; and (6) a minimal synthetic terminator from the rabbit beta globin gene, mRBG. The vector VR1255 expressed 137 times greater than an earlier prototype RSV-based vector. For plasmid vector delivery into nonmuscle tissues, a recently synthesized cationic lipid, GAP-DLRIE, was found to greatly enhance the uptake and expression of plasmid DNA by 100-fold when instilled into the mouse lung. The time-course of CAT expression with GAP-DLRIE indicated that peak expression occurs 2-5 days after intranasal administration and expression diminished to about one-third the peak value by day 21. This cationic lipid may be useful for immunization by pulmonary and perhaps other nonmuscle routes.
authors.affiliation: Vical Inc. San Diego, CA 92121, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Norman
authors.forename: P
authors.initials: P
authors.lastname: Hobart
authors.forename: M
authors.initials: M
authors.lastname: Manthorpe
authors.forename: P
authors.initials: P
authors.lastname: Felgner
authors.forename: C
authors.initials: C
authors.lastname: Wheeler
mesh_terms: D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D010957:Plasmids / Q000235:genetics; D011817:Rabbits; D014611:Vaccination / Q000379:methods*; D019444:Vaccines, DNA / Q000008:administration & dosage; D014765:Viral Vaccines / Q000008:administration & dosage
chemical_list: D019444:Vaccines, DNA; D014765:Viral Vaccines