pEPito

Author(s)
Rudolf Haase, Orestis Argyros, Suet-Ping Wong, Richard P. Harbottle, Hans J. Lipps, Manfred Ogris, Terese Magnusson, Maria G. Vizoso Pinto, Juergen Haas, Armin Baiker
Abstract

Background: The episomal replication of the prototype vector pEPI-1 depends on a transcription unit starting from the constitutively expressed Cytomegalovirus immediate early promoter (CMV-IEP) and directed into a 2000 bp long matrix attachment region sequence (MARS) derived from the human beta-interferon gene. The original pEPI-1 vector contains two mammalian transcription units and a total of 305 CpG islands, which are located predominantly within the vector elements necessary for bacterial propagation and known to be counterproductive for persistent long-term transgene expression.

Results: Here, we report the development of a novel vector pEPito, which is derived from the pEPI-1 plasmid replicon but has considerably improved efficacy both in vitro and in vivo. The pEPito vector is significantly reduced in size, contains only one transcription unit and 60% less CpG motives in comparison to pEPI-1. It exhibits major advantages compared to the original pEPI-1 plasmid, including higher transgene expression levels and increased colony-forming efficiencies in vitro, as well as more persistent transgene expression profiles in vivo. The performance of pEPito-based vectors was further improved by replacing the CMV-IEP with the human CMV enhancer/human elongation factor 1 alpha promoter (hCMV/EF1P) element that is known to be less affected by epigenetic silencing events.

Conclusions: The novel vector pEPito can be considered suitable as an improved vector for biotechnological applications in vitro and for non-viral gene delivery in vivo.

Organisation(s)
External organisation(s)
Ludwig-Maximilians-Universität München, Universität Witten/Herdecke, Imperial College London, University of Edinburgh
Journal
BMC Biotechnology
Volume
10
No. of pages
14
DOI
https://doi.org/10.1186/1472-6750-10-20
Publication date
03-2010
Peer reviewed
Yes
Austrian Fields of Science 2012
301207 Pharmaceutical chemistry
Keywords
Portal url
https://ucris.univie.ac.at/portal/en/publications/pepito(fcb7501e-e1ff-4475-b089-057e8925b437).html