Assessment of Integration-defective HIV-1 and EIAV Vectors In Vitro and In Vivo
The interest in integrase-defective lentiviral vectors (IDLVs) stems from their potential advantage of large cloning capacity and broad cell tropism while avoiding the possibility of insertional mutagenesis. Here, we directly compared the transducing potential of IDLVs based on the equine infectious...
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doaj-4750e5f4e14146d5b05c0ccf43bf2cb42020-11-25T00:32:54ZengElsevierMolecular Therapy: Nucleic Acids2162-25312012-01-011C10.1038/mtna.2012.53Assessment of Integration-defective HIV-1 and EIAV Vectors In Vitro and In VivoScott Ellis0Liang Fong-Wong1Sharifah Iqball2Vinay Thoree3Kyriacos A Mitrophanous4Katie Binley5Oxford BioMedica (UK) Ltd, Medawar Centre, Oxford Science Park, Oxford, UKHenry Wellcome L.I.N.E, University of Bristol, Bristol, UKOxford BioMedica (UK) Ltd, Medawar Centre, Oxford Science Park, Oxford, UKOxford BioMedica (UK) Ltd, Medawar Centre, Oxford Science Park, Oxford, UKOxford BioMedica (UK) Ltd, Medawar Centre, Oxford Science Park, Oxford, UKOxford BioMedica (UK) Ltd, Medawar Centre, Oxford Science Park, Oxford, UKThe interest in integrase-defective lentiviral vectors (IDLVs) stems from their potential advantage of large cloning capacity and broad cell tropism while avoiding the possibility of insertional mutagenesis. Here, we directly compared the transducing potential of IDLVs based on the equine infectious anemia virus (EIAV) to the more commonly described HIV-1 IDLVs. IDLVs were constructed by introducing equivalent single/triple mutations into the integrase catalytic triad. We show that both the single and the triple mutant HIV-1 IDLVs transduce the PC12 cells, but not the C2C12 cells, with similar efficiency to their parental HIV-1 vector. In contrast, the single and triple EIAV IDLVs did not efficiently transduce either differentiated cell line. Moreover, this HIV-1 IDLV-mediated expression was independent of any residual integration activity because reporter expression was lost when cell cycling was restored. Four weeks following stereotactic administration into adult rat brains, only the single HIV-1 IDLV mutant displayed a comparable transduction profile to the parental HIV-1 vector. In contrast, neither EIAV IDLV mutants showed significant reporter gene expression. This work indicates that the transducing potential of IDLVs appears to depend not only on the choice of integrase mutation and type of target cell, but also on the nature of the lentiviral vector.http://www.sciencedirect.com/science/article/pii/S2162253116301196gene therapyintegration-defectivelentiviral vectors |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Scott Ellis Liang Fong-Wong Sharifah Iqball Vinay Thoree Kyriacos A Mitrophanous Katie Binley |
spellingShingle |
Scott Ellis Liang Fong-Wong Sharifah Iqball Vinay Thoree Kyriacos A Mitrophanous Katie Binley Assessment of Integration-defective HIV-1 and EIAV Vectors In Vitro and In Vivo Molecular Therapy: Nucleic Acids gene therapy integration-defective lentiviral vectors |
author_facet |
Scott Ellis Liang Fong-Wong Sharifah Iqball Vinay Thoree Kyriacos A Mitrophanous Katie Binley |
author_sort |
Scott Ellis |
title |
Assessment of Integration-defective HIV-1 and EIAV Vectors In Vitro and In Vivo |
title_short |
Assessment of Integration-defective HIV-1 and EIAV Vectors In Vitro and In Vivo |
title_full |
Assessment of Integration-defective HIV-1 and EIAV Vectors In Vitro and In Vivo |
title_fullStr |
Assessment of Integration-defective HIV-1 and EIAV Vectors In Vitro and In Vivo |
title_full_unstemmed |
Assessment of Integration-defective HIV-1 and EIAV Vectors In Vitro and In Vivo |
title_sort |
assessment of integration-defective hiv-1 and eiav vectors in vitro and in vivo |
publisher |
Elsevier |
series |
Molecular Therapy: Nucleic Acids |
issn |
2162-2531 |
publishDate |
2012-01-01 |
description |
The interest in integrase-defective lentiviral vectors (IDLVs) stems from their potential advantage of large cloning capacity and broad cell tropism while avoiding the possibility of insertional mutagenesis. Here, we directly compared the transducing potential of IDLVs based on the equine infectious anemia virus (EIAV) to the more commonly described HIV-1 IDLVs. IDLVs were constructed by introducing equivalent single/triple mutations into the integrase catalytic triad. We show that both the single and the triple mutant HIV-1 IDLVs transduce the PC12 cells, but not the C2C12 cells, with similar efficiency to their parental HIV-1 vector. In contrast, the single and triple EIAV IDLVs did not efficiently transduce either differentiated cell line. Moreover, this HIV-1 IDLV-mediated expression was independent of any residual integration activity because reporter expression was lost when cell cycling was restored. Four weeks following stereotactic administration into adult rat brains, only the single HIV-1 IDLV mutant displayed a comparable transduction profile to the parental HIV-1 vector. In contrast, neither EIAV IDLV mutants showed significant reporter gene expression. This work indicates that the transducing potential of IDLVs appears to depend not only on the choice of integrase mutation and type of target cell, but also on the nature of the lentiviral vector. |
topic |
gene therapy integration-defective lentiviral vectors |
url |
http://www.sciencedirect.com/science/article/pii/S2162253116301196 |
work_keys_str_mv |
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