PEPito: a Significantly Improved Non-viral Episomal Expression Vector for Mammalian Cells

Overview

Journal BMC Biotechnol

Publisher Biomed Central

Specialty Biotechnology

Date 2010 Mar 17

PMID 20230618

Citations 28

Authors

Rudolf Haase

Orestis Argyros

Suet-Ping Wong

Richard P Harbottle

Hans J Lipps

Manfred Ogris

Terese Magnusson

Maria G Vizoso Pinto

Jurgen Haas

Armin Baiker

Affiliations

Soon will be listed here.

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.

Citing Articles

Impacts of Nucleosome Positioning Elements and Pre-Assembled Chromatin States on Expression and Retention of Transgenes.

Kwizera R, Xie J, Nurse N, Yuan C, Kirchmaier A Genes (Basel). 2024; 15(9).

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VCP/p97 mediates nuclear targeting of non-ER-imported prion protein to maintain proteostasis.

Banik P, Ray K, Kamps J, Chen Q, Luesch H, Winklhofer K Life Sci Alliance. 2024; 7(6).

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Non-Viral Episomal Vector Mediates Efficient Gene Transfer of the β-Globin Gene into K562 and Human Haematopoietic Progenitor Cells.

Lazaris V, Simantirakis E, Stavrou E, Verras M, Sgourou A, Keramida M Genes (Basel). 2023; 14(9).

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Current strategies employed in the manipulation of gene expression for clinical purposes.

Tsai H, Pietrobon V, Peng M, Wang S, Zhao L, Marincola F J Transl Med. 2022; 20(1):535.

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Episomes and Transposases-Utilities to Maintain Transgene Expression from Nonviral Vectors.

Kreppel F, Hagedorn C Genes (Basel). 2022; 13(10).

PMID: 36292757 PMC: 9601623. DOI: 10.3390/genes13101872.

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