The IS2 Element Improves Transcription Efficiency of Integration-Deficient Lentiviral Vector Episomes
| Autor: | Per Anderson, Francisco Martin, Karim Benabdellah, Sabina Sánchez-Hernández, Laura Sánchez, Alejandra Gutierrez-Guerrero, Ana Belén Carrillo-Gálvez, Sandra Rodriguez-Perales, Jesús Chato-Astrain, Jose L. Garcia-Perez, Rocio Martin-Guerra, María Tristán-Manzano, Marina Cortijo-Gutiérrez, Pedro J. Real, Rosa Montes, Ricardo Fernández-Valadés |
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| Přispěvatelé: | Instituto de Salud Carlos III, Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF), Ministerio de Ciencia e Innovación (España), European Research Council |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Cell type HS4 insulator Transgene Genetic enhancement lcsh:RM1-950 Biology Scaffold or matrix attachment regions Viral vector Cell biology 03 medical and health sciences lcsh:Therapeutics. Pharmacology 030104 developmental biology Histone Gene therapy Cell culture Drug Discovery biology.protein Molecular Medicine IDLV Lentiviral vector Progenitor cell Induced pluripotent stem cell |
| Zdroj: | Repisalud Instituto de Salud Carlos III (ISCIII) Molecular Therapy: Nucleic Acids, Vol 13, Iss, Pp 16-28 (2018) |
| Popis: | Integration-defective lentiviral vectors (IDLVs) have become an important alternative tool for gene therapy applications and basic research. Unfortunately, IDLVs show lower transgene expression as compared to their integrating counterparts. In this study, we aimed to improve the expression levels of IDLVs by inserting the IS2 element, which harbors SARs and HS4 sequences, into their LTRs (SE-IS2-IDLVs). Contrary to our expectations, the presence of the IS2 element did not abrogate epigenetic silencing by histone deacetylases. In addition, the IS2 element reduced episome levels in IDLV-transduced cells. Interestingly, despite these negative effects, SE-IS2-IDLVs outperformed SE-IDLVs in terms of percentage and expression levels of the transgene in several cell lines, including neurons, neuronal progenitor cells, and induced pluripotent stem cells. We estimated that the IS2 element enhances the transcriptional activity of IDLV LTR circles 6- to 7-fold. The final effect the IS2 element in IDLVs will greatly depend on the target cell and the balance between the negative versus the positive effects of the IS2 element in each cell type. The better performance of SE-IS2-IDLVs was not due to improved stability or differences in the proportions of 1-LTR versus 2-LTR circles but probably to a re-positioning of IS2-episomes into transcriptionally active regions. This study is financed by the ISCIII Health Research Fund (Spain)and the European Regional Development Fund (FEDER) throughresearch grants PI12/01097 and PI15/02015 (F.M.) and CD09/0020,PI15/00794, and CPII15/00032 (P.A.); the ISCIII Cellular TherapyNetwork (TerCel, RD12/0019/0006) (F.M.); a Juan de la Cierva fellowship (JCI_2012_12666; to R.M.); the CICE and CS of the Juntade Andalucía FEDER/European Cohesion Fund (FSE) for Andalucía2007-2013 through research grants P09-CTS-04532, PI-57069, PI-0001/2009, and PAIDI-Bio-326 (F.M.) and PI-0160/2012 and PI-0014-2016 (K.B.);the Ministerio de Economía, Industria y competi-tividad through research grant SAF2017-89745-R (J.L.G.-P.); aRYC contract. (RYC-2015-18382; to P/J.R.); an FPU fellowship(FPU16/05467; to M.T.-M.); a PEJ contract (PEJ-2014-A-46314; to A.B.C.-G. and PEJ-2014-A-17105 S.S.-H.); the European Research (ERC-Consolidator ERC-STG-2012-233764); and a privatedonation from Ms. Francisca Serrano (Trading y Bolsa para Torpes,Granada, Spain).We also wish to tand a private donation from Ms. Francisca Serrano (Trading y Bolsa para Torpes, Granada, Spain).We also wish to thank Michael O’Shea for proofreading the article. J.L.G.-P’s lab is supported by MINECO-FEDER (SAF2017-89745-R). Sí |
| Databáze: | OpenAIRE |
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