RT Journal Article
T1 The IS2 Element Improves Transcription Efficiency of Integration-Deficient Lentiviral Vector Episomes.
A1 Sánchez-Hernández, Sabina
A1 Gutierrez-Guerrero, Alejandra
A1 Martín-Guerra, Rocío
A1 Cortijo-Gutierrez, Marina
A1 Tristán-Manzano, María
A1 Rodriguez-Perales, Sandra
A1 Sanchez, Laura
A1 Garcia-Perez, Jose Luis
A1 Chato-Astrain, Jesus
A1 Fernandez-Valades, Ricardo
A1 Carrillo-Galvez, Ana Belén
A1 Anderson, Per
A1 Montes, Rosa
A1 Real, Pedro J
A1 Martin, Francisco
A1 Benabdellah, Karim
K1 HS4 insulator
K1 IDLV
K1 gene therapy
K1 lentiviral vector
K1 scaffold or matrix attachment regions
AB 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.
SN 2162-2531
YR 2018
FD 2018-08-18
LK http://hdl.handle.net/10668/12961
UL http://hdl.handle.net/10668/12961
LA en
DS RISalud
RD Mar 14, 2025