The extragenic spacer length between the 5' and 3' ends of the transgene expression cassette affects transgene silencing from plasmid-based vectors.

Molecular Therapy : the Journal of the American Society of Gene Therapy
Jiamiao LuM A Kay

Abstract

In quiescent tissues, minicircle DNA vectors provide at least 10 times higher sustained levels of transgene expression compared to that achieved with a canonical plasmid containing the same expression cassette. It is not known if there is a specific DNA sequence or structure that is needed for DNA silencing. To directly address this question, we substituted the bacterial plasmid DNA with various lengths of extragenic spacer DNAs between the 5' and 3' ends of the transgene expression cassette and determined the expression profiles using two different reporter expression cassettes. Both the human alphoid repeat (AR) and randomly generated DNA sequences of ≥1 kb in length resulted in transgene silencing while shorter spacers, ≤500 bp exhibited similar transgene expression patterns to conventional minicircle DNA vectors. In contrast, when the ≥1 kb random DNA (RD) sequences were expressed as part of the 3'-untranslated region (UTR) transgene silencing was not observed. These data suggest that the length and not the sequence or origin of the extragenic DNA flanking the expression cassette is responsible for plasmid-mediated transgene silencing. This has implications for the design of nonviral vectors for gene transfer applications a...Continue Reading

References

Feb 24, 1998·Gene Therapy·A M DarquetJ Crouzet
Oct 19, 2001·Biochemical and Biophysical Research Communications·E A GrzybowskaJ A Siedlecki
Aug 30, 2003·Molecular Therapy : the Journal of the American Society of Gene Therapy·Zhi-Ying ChenMark A Kay
Aug 18, 2004·Nature Genetics·Justin M O'SullivanNick J Proudfoot
Dec 2, 2005·Genes & Development·Athar Ansari, Michael Hampsey
Apr 26, 2007·Molecular Therapy : the Journal of the American Society of Gene Therapy·Efren RiuMark A Kay
Feb 7, 2008·Molecular Therapy : the Journal of the American Society of Gene Therapy·Zhi-Ying ChenMark A Kay
Sep 4, 2008·The Journal of Gene Medicine·Peter MayrhoferWolfgang Jechlinger
Feb 11, 2009·Nature Reviews. Genetics·Cizhong Jiang, B Franklin Pugh
Nov 26, 2010·Nature Biotechnology·Mark A KayZhi-Ying Chen

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Citations

Jul 9, 2013·Trends in Biotechnology·Pedro H Oliveira, Juergen Mairhofer
Mar 6, 2013·Molecular Therapy : the Journal of the American Society of Gene Therapy·Jiamiao LuMark A Kay
May 22, 2013·Molecular Therapy : the Journal of the American Society of Gene Therapy·Wynette M DietzMark J Osborn
Jan 30, 2014·International Journal of Pharmaceutics·Genki N KandaHiroyuki Kamiya
Mar 27, 2016·Journal of Chromatography. a·Cláudia P A AlvesDuarte Miguel F Prazeres
Mar 4, 2014·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Hiu Man ViecelliBeat Thöny
May 6, 2016·Applied Microbiology and Biotechnology·Michaela ŠimčíkováGabriel A Monteiro
May 11, 2016·Biotechnology & Genetic Engineering Reviews·Michaela ŠimčíkováGabriel A Monteiro
Apr 30, 2016·Gene Therapy·P N MatkarK K Singh
Jul 1, 2016·Molecular Therapy : the Journal of the American Society of Gene Therapy·Xiaoou JiangVolker Patzel
Feb 13, 2014·New Biotechnology·Jeff Jia Cheng HouTrent P Munro
Feb 18, 2017·Genes·Cinnamon L HardeeLynn Zechiedrich
Jun 13, 2017·Human Vaccines & Immunotherapeutics·John J SuschakConnie S Schmaljohn
Apr 28, 2018·Biotechnology Journal·Xue WangMasamichi Kamihira
Aug 23, 2017·Human Gene Therapy·Ana Krhac LevacicErnst Wagner
Sep 4, 2018·Biological & Pharmaceutical Bulletin·Tetsuya SuzukiHiroyuki Kamiya
Sep 18, 2014·The Korean Journal of Internal Medicine·Sebastian DieckeJi Hyeon Ju
Jan 1, 2014·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Charles H JonesBlaine A Pfeifer
Apr 25, 2020·PloS One·Dina R SafinaSergey V Kostrov
Apr 17, 2020·Molecular Therapy. Methods & Clinical Development·John J SuschakConnie S Schmaljohn
Feb 12, 2017·Pharmacology & Therapeutics·Christopher D ZahmDouglas G McNeel
Apr 20, 2021·Molecular Therapy. Methods & Clinical Development·Marie SoukupováMichele Simonato
Jun 25, 2015·Microbiology Spectrum·Duarte Miguel F Prazeres, Gabriel A Monteiro
Oct 8, 2013·Molecular Pharmaceutics·Charles H JonesBlaine A Pfeifer

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