Re-structuring lentiviral vectors to express genomic RNA via cap-dependent translation.


Journal

Molecular therapy. Methods & clinical development
ISSN: 2329-0501
Titre abrégé: Mol Ther Methods Clin Dev
Pays: United States
ID NLM: 101624857

Informations de publication

Date de publication:
12 Mar 2021
Historique:
received: 06 03 2020
accepted: 01 12 2020
entrez: 8 2 2021
pubmed: 9 2 2021
medline: 9 2 2021
Statut: epublish

Résumé

Lentiviral (LV) vectors based on human immunodeficiency virus type I (HIV-1) package two copies of their single-stranded RNA into vector particles. Normally, this RNA genome is reverse transcribed into a double-stranded DNA provirus that integrates into the cell genome, providing permanent gene transfer and long-term expression. Integration-deficient LV vectors have been developed to reduce the frequency of genomic integration and thereby limit their persistence in dividing cells. Here, we describe optimization of a reverse-transcriptase-deficient LV vector, which enables direct translation of LV RNA genomes upon cell entry, for transient expression of vector payloads as mRNA without a DNA intermediate. We have engineered a novel LV genome arrangement in which HIV-1 sequences are removed from the 5' end, to enable ribosomal entry from the 5' 7-methylguanylate cap for efficient translation of the vector payload. We have shown that this LV-mediated mRNA delivery platform provides transient transgene expression

Identifiants

pubmed: 33553484
doi: 10.1016/j.omtm.2020.12.005
pii: S2329-0501(20)30253-9
pmc: PMC7838728
doi:

Types de publication

Journal Article

Langues

eng

Pagination

357-365

Subventions

Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 210774/Z/18/Z
Pays : United Kingdom

Informations de copyright

© 2020 The Authors.

Déclaration de conflit d'intérêts

J.R.C. and G.D.B. are inventors on a patent application covering the technology described in this manuscript. The other authors declare no competing interests.

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Auteurs

John R Counsell (JR)

Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Programme, UCL Great Ormond Street Institute of Child Health, 30 Guilford Street, London, UK.
NIHR Great Ormond Street Hospital Biomedical Research Centre, 30 Guilford Street, London WC1N 1EH, UK.

Guillaume De Brabandere (G)

Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Programme, UCL Great Ormond Street Institute of Child Health, 30 Guilford Street, London, UK.
NIHR Great Ormond Street Hospital Biomedical Research Centre, 30 Guilford Street, London WC1N 1EH, UK.

Rajvinder Karda (R)

Gene Transfer Technology Group, Institute for Women's Health, University College London, 86-96 Chenies Mews, London, UK.

Marc Moore (M)

Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Programme, UCL Great Ormond Street Institute of Child Health, 30 Guilford Street, London, UK.
NIHR Great Ormond Street Hospital Biomedical Research Centre, 30 Guilford Street, London WC1N 1EH, UK.

Antonio Greco (A)

Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Programme, UCL Great Ormond Street Institute of Child Health, 30 Guilford Street, London, UK.
NIHR Great Ormond Street Hospital Biomedical Research Centre, 30 Guilford Street, London WC1N 1EH, UK.

Alysha Bray (A)

Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Programme, UCL Great Ormond Street Institute of Child Health, 30 Guilford Street, London, UK.
NIHR Great Ormond Street Hospital Biomedical Research Centre, 30 Guilford Street, London WC1N 1EH, UK.

Juan Antinao Diaz (JA)

Gene Transfer Technology Group, Institute for Women's Health, University College London, 86-96 Chenies Mews, London, UK.

Dany P Perocheau (DP)

Gene Transfer Technology Group, Institute for Women's Health, University College London, 86-96 Chenies Mews, London, UK.

Ulrike Mock (U)

NIHR Great Ormond Street Hospital Biomedical Research Centre, 30 Guilford Street, London WC1N 1EH, UK.

Simon N Waddington (SN)

Gene Transfer Technology Group, Institute for Women's Health, University College London, 86-96 Chenies Mews, London, UK.
MRC Antiviral Gene Therapy Research Unit, Faculty of Health Sciences, University of the Witswatersrand, Johannesburg, South Africa.

Classifications MeSH