pCOR: a new design of plasmid vectors for nonviral gene therapy

Abstract

A totally redesigned host/vector system with improved initiator protein, protein, encoded by the pir gene limiting properties in terms of safety has been developed. The its host range to bacterial strains that produce this transpCOR plasmids are narrow-host range plasmid vectors for acting protein; (2) the plasmid's selectable marker is not an nonviral gene therapy. These plasmids contain a conantibiotic resistance gene but a gene encoding a bacterial ditional origin of replication and must be propagated in a suppressor tRNA. Optimized E. coli hosts supporting specifically engineered E. coli host strain, greatly reducing pCOR replication and selection were constructed. High the potential for propagation in the environment or in yields of supercoiled pCOR monomers were obtained (100 treated patients. The pCOR backbone has several features mg/l) through fed-batch fermentation. pCOR vectors carrythat increase safety in terms of dissemination and selecing the luciferase reporter gene gave high levels of lucifertion: (1) the origin of replication requires a plasmid-specific ase activity when injected into murine skeletal muscle.

Electrotransfer of naked DNA in the skeletal muscles of animal models of muscular dystrophies

“…65 The plasmid preparation contained a high percentage of supercoiled DNA (99.9%), and no RNA was detectable by gel electrophoresis. The plasmid size was 5 kb.…”

Non-viral gene transfer of murine spleen cells achieved by in vivo electroporation

“…The plasmid pXL 3031 encoding luciferase (pCMV-Luc) contained the cytomegalovirus (CMV) promoter (nucleotides 229-890 of pcDNA3, Invitrogen) inserted upstream of the coding sequence of the modified cytosolic luc+ luciferase. 30 The pCMV-empty plasmid was obtained from the pCMV-Luc in which the luciferase sequence was excised. The pCMV-SeAP plasmid (pRVX-SeAP2) was obtained by cloning the human SeAP (pSEAP2-Control plasmid; Clontech) downstream of the pCMV in the pcDNA3.1 backbone (Invitrogen).…”

An improved anion-exchange HPLC method for the detection and purification of adenoviral particles

“…18 The technology was simplified by replacing the ColE1 origin of replication by the ori ␥ of R6K in the suicide shuttle 19 which allows recombination in any recA + E. coli strain. Plasmid pXL3185 is 57.6 kbp long and derives from the very stable RK2 vector.…”