Replicative minicircle vectors with improved expression

Abstract

The present invention relates to the production and use of covalently closed circular (ccc) recombinant DNA molecules such as plasmids, cosmids, bacterial artificial chromosomes (BACs), bacteriophages, viral vectors and hybrids thereof, and more particularly to vector modifications that improve expression of said DNA molecules.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application claims priority to U.S. Provisional Patent Application Ser. No. 61 / 796,765, entitled “Replicative Minicircle vectors with improved expression” which was filed Nov. 19, 2012, the entire contents of which are incorporated herein by reference and to U.S. Provisional Patent Application Ser. No. 61 / 743,219, entitled “DNA Plasmids With Improved Expression” which was filed Aug. 29, 2012, the entire contents of which are incorporated herein by reference.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002]This invention was supported in part with government support under Grant No. R44GM080768, awarded by the National Institutes of Health. The government has certain rights in this invention.FIELD OF THE INVENTION[0003]The present invention relates to a family of eukaryotic expression plasmids useful for gene therapy, obtaining improved genetic immunization, natural interferon production, and more particularly, for ...

AI Technical Summary

Benefits of technology

[0016]Another object of the invention is to provide eukaryotic expression vectors containing short spacer regions that may be efficiently manufactured.

Problems solved by technology

The art teaches that one of the limitations of application of plasmid therapies and plasmid vaccines is regulatory agency (e.g. Food and Drug Administration, European Medicines Agency) safety concerns regarding 1) plasmid transfer and replication in endogenous bacterial flora, or 2) plasmid encoded selection marker expression in human cells, or endogenous bacterial flora.

The art further teaches that one of the limitations of application of plasmid therapies and vaccines is that transgene expression is generally very low.

This results in short duration transgene expression.

However, methods to manufacture MIDGE and minicircle vectors are expensive and not easily scalable.

Creating terminal loops on MIDGE vectors in vitro is problematic, requiring in vitro ligation of annealed primers to restriction digested vector.

These production procedures are very inefficient.

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