+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ PDF Full Text
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Translate
+ Recently Requested

Physiological levels of HBB transgene expression from S/MAR element-based replicating episomal vectors

Physiological levels of HBB transgene expression from S/MAR element-based replicating episomal vectors

Journal of Biotechnology 143(2): 85-94

Replicating episomal vectors (REV) are in principle able to provide long-term transgene expression in the absence of integration into the target cell genome. The scaffold/matrix attachment region (S/MAR) located 5' of the human beta-interferon gene (IFNB1) has been shown to confer a stable episomal replication and retention function within plasmid vectors when stably transfected and selected in mammalian cells. The minimal requirement for the IFNB1 S/MAR to function in DNA replication and episomal retention is transcription through this element. We used the erythroid beta-globin locus control region-beta-globin gene (betaLCR-HBB) microlocus cassette as a model to assess tissue-specific expression from within an IFNB1 S/MAR-based plasmid REV. The betaLCR-HBB plus S/MAR combination constructs provided either high or low levels of transcription through the S/MAR element. Our results show that the betaLCR-HBB microlocus is able to reproducibly and stably express at full physiological levels on an episome copy number basis. In addition, our data show that even low levels of transcription from betaLCR-HBB through the S/MAR element are sufficient to allow efficient episomal replication and retention. These data provide the principles upon which generic and flexible expression cassette-S/MAR-based REVs can be designed for a wide range of applications.

(PDF emailed within 0-6 h: $19.90)

Accession: 055001978

Download citation: RISBibTeXText

PMID: 19559736

DOI: 10.1016/j.jbiotec.2009.06.018

Related references

Self-replicating episomal expression vectors conferring tissue-specific gene expression. Official Gazette of the United States Patent & Trademark Office Patents 1286(4), Sep 28, 2004

Ectopic expression of AID in a non-B cell line triggers A:T and G:C point mutations in non-replicating episomal vectors. Plos One 3(1): E1480-E1480, 2008

LCR-mediated, long-term tissue-specific gene expression within replicating episomal plasmid and cosmid vectors. Gene Therapy 9(5): 327-336, 2002

Long-term episomal transgene expression from mitotically stable integration-deficient lentiviral vectors. Human Gene Therapy 25(5): 428-442, 2014

Conditionally replicating lentiviral-hybrid episomal vectors for suicide gene therapy. Antiviral Research 80(3): 288-294, 2008

Barriers for Deriving Transgene-Free Pig iPS Cells with Episomal Vectors. Stem Cells 33(11): 3228-3238, 2016

Expression in human cells of exogenous genes using episomal vectors based on bk virus dna. Igiene Moderna 89(1): 156-162, 1988

Generation of transgene-free iPSC lines from human normal and neoplastic blood cells using episomal vectors. Methods in Molecular Biology 997: 163-176, 2013

Transfection of primary human keratinocytes with functional gene constructs in ebv based episomal expression vectors. Journal of Cellular Biochemistry Supplement (14 PART A): 365, 1990

Woodchuck hepatitis virus post-transcriptional regulation element enhances transgene expression from adenovirus vectors. Biochimica et Biophysica Acta 1621(3): 266-271, 11 June, 2003

Effects of viral strain, transgene position, and target cell type on replication kinetics, genomic stability, and transgene expression of replication-competent murine leukemia virus-based vectors. Journal of Virology 81(13): 6973-6983, 2007

Humoral and cellular immune responses to HIV-1 Nef in mice DNA-immunised with non-replicating or self-replicating expression vectors. Vaccine 18(5-6): 460-467, 1999

Incorporation of a SAR element into retroviral gene transfer vectors leads to an orientation dependent increase in transgene expression in stem cells. Blood 98(11 Part 1): 746a, November 16, 2001

Enhancing Transgene Expression from Recombinant AAV8 Vectors in Different Tissues Using Woodchuck Hepatitis Virus Post-Transcriptional Regulatory Element. International Journal of Medical Sciences 13(4): 286-291, 2016