The IS2 Element Improves Transcription Efficiency of Integration-Deficient Lentiviral Vector Episomes
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...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2018-12-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253118302129 |
| _version_ | 1818525395516915712 |
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| author | Sabina Sánchez-Hernández Alejandra Gutierrez-Guerrero Rocío Martín-Guerra Marina Cortijo-Gutierrez María Tristán-Manzano Sandra Rodriguez-Perales Laura Sanchez Jose Luis Garcia-Perez Jesus Chato-Astrain Ricardo Fernandez-Valades Ana Belén Carrillo-Galvez Per Anderson Rosa Montes Pedro J. Real Francisco Martin Karim Benabdellah |
| author_facet | Sabina Sánchez-Hernández Alejandra Gutierrez-Guerrero Rocío Martín-Guerra Marina Cortijo-Gutierrez María Tristán-Manzano Sandra Rodriguez-Perales Laura Sanchez Jose Luis Garcia-Perez Jesus Chato-Astrain Ricardo Fernandez-Valades Ana Belén Carrillo-Galvez Per Anderson Rosa Montes Pedro J. Real Francisco Martin Karim Benabdellah |
| author_sort | Sabina Sánchez-Hernández |
| collection | DOAJ |
| description | 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. Keywords: IDLV, HS4 insulator, gene therapy, scaffold or matrix attachment regions, lentiviral vector |
| first_indexed | 2024-12-11T06:08:47Z |
| format | Article |
| id | doaj.art-72961106e0f041648c1d03824c09a7c3 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-11T06:08:47Z |
| publishDate | 2018-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-72961106e0f041648c1d03824c09a7c32022-12-22T01:18:11ZengElsevierMolecular Therapy: Nucleic Acids2162-25312018-12-01131628The IS2 Element Improves Transcription Efficiency of Integration-Deficient Lentiviral Vector EpisomesSabina Sánchez-Hernández0Alejandra Gutierrez-Guerrero1Rocío Martín-Guerra2Marina Cortijo-Gutierrez3María Tristán-Manzano4Sandra Rodriguez-Perales5Laura Sanchez6Jose Luis Garcia-Perez7Jesus Chato-Astrain8Ricardo Fernandez-Valades9Ana Belén Carrillo-Galvez10Per Anderson11Rosa Montes12Pedro J. Real13Francisco Martin14Karim Benabdellah15Genomic Medicine Department, GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, SpainGenomic Medicine Department, GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, SpainGenomic Medicine Department, GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, SpainGenomic Medicine Department, GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, SpainGenomic Medicine Department, GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, SpainMolecular Cytogenetics and Genome Editing Unit, Human Cancer Genetics Department, CNIO, Melchor Fernandez Almagro 3, 28029 Madrid, SpainGenomic Medicine Department, GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, SpainGenomic Medicine Department, GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, SpainDepartment of Histology, Tissue Engineering Group, University of Granada, Granada, SpainPediatric Surgery Department, University Hospital “Virgen de las Nieves,” Avda. Fuerzas Armadas 2, 18014 Granada, SpainOncology Department, GENYO, Centre for Genomics and Oncology, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, SpainLentiStem Biotech, GENYO, Avda. de la Ilustración 114, 18016 PTS Granada, Spain; Oncology Department, GENYO, Centre for Genomics and Oncology, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, SpainOncology Department, GENYO, Centre for Genomics and Oncology, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, SpainOncology Department, GENYO, Centre for Genomics and Oncology, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, Spain; Departament of Biochemistry and Molecular Biology I, University of Granada, Granada, SpainGenomic Medicine Department, GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, Spain; LentiStem Biotech, GENYO, Avda. de la Ilustración 114, 18016 PTS Granada, Spain; Corresponding author: Francisco Martin, Genomic Medicine Department, GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, Health Sciences Technology Park, Avda. de la Ilustración 114, 18016 Granada, SpainGenomic Medicine Department, GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, 18016 Granada, Spain; LentiStem Biotech, GENYO, Avda. de la Ilustración 114, 18016 PTS Granada, Spain; Corresponding author: Karim Benabdellah, Genomic Medicine Department, GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Government, PTS Granada, Health Sciences Technology Park, Avda. de la Ilustración 114, 18016 Granada, SpainIntegration-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. Keywords: IDLV, HS4 insulator, gene therapy, scaffold or matrix attachment regions, lentiviral vectorhttp://www.sciencedirect.com/science/article/pii/S2162253118302129 |
| spellingShingle | Sabina Sánchez-Hernández Alejandra Gutierrez-Guerrero Rocío Martín-Guerra Marina Cortijo-Gutierrez María Tristán-Manzano Sandra Rodriguez-Perales Laura Sanchez Jose Luis Garcia-Perez Jesus Chato-Astrain Ricardo Fernandez-Valades Ana Belén Carrillo-Galvez Per Anderson Rosa Montes Pedro J. Real Francisco Martin Karim Benabdellah The IS2 Element Improves Transcription Efficiency of Integration-Deficient Lentiviral Vector Episomes Molecular Therapy: Nucleic Acids |
| title | The IS2 Element Improves Transcription Efficiency of Integration-Deficient Lentiviral Vector Episomes |
| title_full | The IS2 Element Improves Transcription Efficiency of Integration-Deficient Lentiviral Vector Episomes |
| title_fullStr | The IS2 Element Improves Transcription Efficiency of Integration-Deficient Lentiviral Vector Episomes |
| title_full_unstemmed | The IS2 Element Improves Transcription Efficiency of Integration-Deficient Lentiviral Vector Episomes |
| title_short | The IS2 Element Improves Transcription Efficiency of Integration-Deficient Lentiviral Vector Episomes |
| title_sort | is2 element improves transcription efficiency of integration deficient lentiviral vector episomes |
| url | http://www.sciencedirect.com/science/article/pii/S2162253118302129 |
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