A rapid and quantitative assay for measuring neighboring gene activation by vector proviruses

Molecular Therapy : the Journal of the American Society of Gene Therapy
Paul C HendrieDavid W Russell

Abstract

A simple, quantitative assay for measuring the oncogenic potential of integrating vectors is needed in order to improve vector design and safety. In this study, we have developed a transient plasmid-based assay to measure the activation of a reporter gene by an adjacent vector provirus. Plasmid pACT contains a luciferase cassette driven by a minimal, enhancerless promoter, into which vector proviruses are inserted upstream for evaluation by luciferase assays and northern blots. In a comparison of analogous vectors based on murine leukemia virus (MLV), human immunodeficiency virus (HIV), and foamy virus (FV), we observed significant enhancer activity and read-through transcription from MLV proviruses, and significant read-through transcription from HIV proviruses. HIV and FV proviruses containing an internal MLV long-terminal repeat (LTR) promoter also had significant enhancer activity, which was not observed with an internal promoter from the murine phosphoglycerate kinase-1 gene, PGK. These results demonstrate that neighboring gene activation can be limited by using internal promoter(s) lacking enhancer activity, especially when present in an FV vector backbone that prevents read-through transcription. Although the pACT assay ...Continue Reading

References

Jul 1, 1977·The Journal of General Virology·F L GrahamR Nairn
Jun 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·M Gossen, H Bujard
Oct 25, 1991·Nucleic Acids Research·M W McBurneyK Jardine
Jul 1, 1989·Genes & Development·N LevittN J Proudfoot
May 16, 1996·Biochimica Et Biophysica Acta·J Jonkers, A Berns
Nov 13, 1998·Proceedings of the National Academy of Sciences of the United States of America·A KrummM Groudine
Dec 10, 1999·Proceedings of the National Academy of Sciences of the United States of America·F Recillas-TargaG Felsenfeld
Apr 28, 2000·Science·M Cavazzana-CalvoA Fischer
Apr 20, 2002·Science·Zhixiong LiChristopher Baum
Jun 20, 2002·Journal of Virology·Anne-Kathrin ZaissLung-Ji Chang
Sep 17, 2002·Molecular Therapy : the Journal of the American Society of Gene Therapy·Grant TrobridgeDavid W Russell
Aug 9, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Mike ThemisCharles Coutelle
Jan 24, 2006·Proceedings of the National Academy of Sciences of the United States of America·Grant D TrobridgeDavid W Russell
Jul 26, 2006·Proceedings of the National Academy of Sciences of the United States of America·Yan ShouBrian P Sorrentino
Sep 9, 2006·The Journal of Gene Medicine·Stephanie LaufsStefan Fruehauf
Sep 14, 2006·Blood·Hans-Peter KiemDavid W Russell

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Citations

Jul 16, 2010·Gene Therapy·O Erlwein, M O McClure
Jun 26, 2008·Molecular Therapy : the Journal of the American Society of Gene Therapy·Jingqiong HuCynthia E Dunbar
Aug 27, 2009·Molecular Therapy : the Journal of the American Society of Gene Therapy·Paritha I ArumugamPunam Malik
Apr 1, 2010·Molecular Therapy : the Journal of the American Society of Gene Therapy·Samantha L GinnIan E Alexander
Oct 28, 2011·Molecular Therapy : the Journal of the American Society of Gene Therapy·David R DeyleDavid W Russell
Mar 28, 2013·Molecular Therapy : the Journal of the American Society of Gene Therapy·Thomas R BauerDennis D Hickstein
Feb 10, 2009·Human Gene Therapy·Grant D TrobridgeHans-Peter Kiem
Aug 27, 2010·Human Gene Therapy·Ken OhmineDavid W Russell
Jul 2, 2010·Journal of Virology·David R DeyleDavid W Russell
Oct 14, 2011·Viruses·Dirk Lindemann, Axel Rethwilm
Dec 12, 2012·Viruses·Grant D TrobridgeHans-Peter Kiem
Oct 25, 2013·Viruses·Miles E Olszko, Grant D Trobridge
Aug 15, 2013·Medical Science Monitor Basic Research·Dimitris ZacharoulisGeorge Vassilopoulos
Jul 24, 2012·Therapeutic Delivery·Otto Erlwein, Myra McClure
Mar 8, 2011·Expert Opinion on Biological Therapy·Grant D Trobridge
Sep 12, 2009·Expert Opinion on Biological Therapy·Grant D Trobridge
Dec 31, 2015·Human Gene Therapy·Diana L BrowningGrant D Trobridge
May 3, 2016·Molecular Therapy : the Journal of the American Society of Gene Therapy·Nathan Paul SweeneyMyra Olga McClure
Sep 1, 2016·Molecular Therapy. Methods & Clinical Development·Elizabeth M EversonGrant D Trobridge
Nov 5, 2016·Scientific Reports·Jonah D HocumGrant D Trobridge
Dec 27, 2016·Gene Therapy·D L BrowningG D Trobridge
Jun 24, 2018·The Journal of Gene Medicine·Elizabeth M EversonGrant D Trobridge
Sep 20, 2008·Gene Therapy·G Vassilopoulos, A Rethwilm
Apr 21, 2017·Biomedicines·Diana L Browning, Grant D Trobridge
Mar 29, 2016·Biomedicines·Arun K Nalla, Grant D Trobridge
Mar 25, 2020·Viruses·Emmanouil SimantirakisGeorge Vassilopoulos

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