Abstract
A wider application of minicircle (MC) vectors in gene therapy research depends critically on the ability to purify supercoiled (sc) MC from related miniplasmid (MP) and parental plasmid (PP) impurities. This protocol describes a purification strategy that combines the in vitro enzymatic relaxation of sc MP and PP impurities by a nicking endonuclease, and topoisomer separation and RNA clearance by hydrophobic interaction chromatography. The time required to follow the full protocol, from production to isolation of sc MC, is approximately 50 h. The process delivers sc MCs that are virtually free from MP, PP, RNA, and protein impurities.
References
Feb 24, 1998·Gene Therapy·A M DarquetJ Crouzet
Aug 6, 1999·Gene Therapy·A M DarquetD Scherman
Dec 12, 2001·Microbiology·A KhlebnikovJ D Keasling
Aug 30, 2003·Molecular Therapy : the Journal of the American Society of Gene Therapy·Zhi-Ying ChenMark A Kay
Feb 5, 2004·Molecular Therapy : the Journal of the American Society of Gene Therapy·Hongmei ZhaoNelson S Yew
Feb 11, 2005·Human Gene Therapy·Zhi-Ying ChenMark A Kay
Apr 14, 2005·Journal of Molecular Biology·Daniel F HeiterGeoffrey G Wilson
Sep 24, 2005·Journal of Molecular Microbiology and Biotechnology·Wolfgang JechlingerPeter Mayrhofer
Dec 8, 2005·DNA and Cell Biology·Irina TumanovaAnthony P Green
Apr 26, 2007·Molecular Therapy : the Journal of the American Society of Gene Therapy·Efren RiuMark A Kay
Mar 11, 2008·Molecular Biotechnology·Juergen Mairhofer, Reingard Grabherr
Sep 4, 2008·The Journal of Gene Medicine·Peter MayrhoferWolfgang Jechlinger
Nov 26, 2010·Nature Biotechnology·Mark A KayZhi-Ying Chen
Apr 7, 2011·Nature Reviews. Genetics·Mark A Kay
May 9, 2012·Molecular Therapy : the Journal of the American Society of Gene Therapy·Jiamiao LuMark A Kay
Sep 28, 2012·Pharmaceutical Biology·Huaben BoShulin Huang
Nov 28, 2012·Molecular Therapy : the Journal of the American Society of Gene Therapy·Lia E Gracey ManiarMark A Kay
May 22, 2013·Molecular Therapy : the Journal of the American Society of Gene Therapy·Wynette M DietzMark J Osborn
Sep 3, 2013·Journal of Pharmaceutical Sciences·Jonathan De La VegaDuarte Miguel F Prazeres
Oct 16, 2015·Nature·Luigi Naldini
Mar 16, 2016·Scientific Reports·Mustafa M MunyeStephen L Hart
Mar 27, 2016·Journal of Chromatography. a·Cláudia P A AlvesDuarte Miguel F Prazeres
May 6, 2016·Applied Microbiology and Biotechnology·Michaela ŠimčíkováGabriel A Monteiro
Feb 18, 2017·Genes·Cinnamon L HardeeLynn Zechiedrich
Citations
Sep 12, 2018·Human Gene Therapy·Joana SerraCláudia L da Silva
Apr 20, 2021·The Journal of Gene Medicine·Joana SerraDuarte Miguel F Prazeres
Jun 13, 2021·Analytical Biochemistry·Cláudia P A AlvesGabriel A Monteiro