TABLE 67.1.

Some Major Approaches^a in Plasmid Vaccine Optimization

Plasmid Modification	Sequence Modifications	Gene Adjuvants and Formulations	Delivery Enhancement
•Promoter choice	•Modification of GC/AT content	•Molecular adjuvants	•Electroporation
•Backbone size	•Species codon optimization	•Cytokines	•Jet injector
•Enhancer elements	•RNA optimizations	•Chemokines	•Gene gun
•Transactivation sequences	•Strong Kozack start sequence	•Toll-like receptors	•Skin abrasion
•Internal termination sequence	•Leader sequence	•HSP	•Microneedle
•Poly AAA tract	•Termination sequence	•Costimulatory genes	•Topical patch
•Optimized ORI for production	•Localization sequences	•Transcription factors	•Needle-free systems
•Antibiotic selection sequence for stable production	•Glycosylation sequences	•Adhesion molecules	•Hydrodynamic delivery
	•Immunogen sequence optimization	•Formulations
	•Epitope strings	•Alum
	•Consensus	•Saponin
	•Mosaic	•Nanoparticles
	•Center-of-tree	•Liposomes
	•Matrix immunogens	•Polymers
	•Polyvalency or particle formation
	•Localization sequences
	•Designer immunogens

HSP, heat shock proteins; ORI, origin of replication (site where DNA replication is initiated).

^aAlternative approaches to modification of plasmids, coding and noncoding sequence changes, formulation or adjuvanting, and delivery methods that can improve insert expression or immunogenicity are indicated.