Titel
Inhibition of adenovirus replication by CRISPR-Cas9-mediated targeting of the viral E1A gene
Autor*in
Zrinka Didara
Department of Life Sciences, University of Applied Sciences Krems
Florian Reithofer
Department of Life Sciences, University of Applied Sciences Krems
Karina Zöttl
Department of Life Sciences, University of Applied Sciences Krems
... show all
Abstract
DNA-targeting CRISPR-Cas systems are able to cleave dsDNA in mammalian cells. Accordingly, they have been employed to target the genomes of dsDNA viruses, mostly when present in cells in a non-replicative state with low copy numbers. However, the sheer amount of viral DNA produced within a very short time by certain lytically replicating viruses potentially brings the capacities of CRISPR-Cas systems to their limits. The accessibility of viral DNA replication sites, short time of accessibility of the DNA before encapsidation, or its complexation with shielding proteins are further potential hurdles. Adenoviruses are fast-replicating dsDNA viruses for which no approved antiviral therapy currently exists. We evaluated the potency of CRISPR-Cas9 in inhibiting the replication of human adenovirus 5 in vitro by targeting its master regulator E1A with a set of guide RNAs and observed a decrease in infectious virus particles by up to three orders of magnitude. Target DNA cleavage also negatively impacted the amount of viral DNA accumulated during the infection cycle. This outcome was mainly caused by specific deletions, inversions, and duplications occurring between target sites, which abolished most E1A functions in most cases. Additionally, we compared two strategies for multiplex gRNA expression and obtained comparable results.
Stichwort
MT: RNA/DNA editingCRISPR-Cas9adenovirusinfectionvirustherapy
Objekt-Typ
Sprache
Englisch [eng]
Persistent identifier
phaidra.univie.ac.at/o:2068845
Erschienen in
Titel
Molecular Therapy - Nucleic Acids
Band
32
ISSN
2162-2531
Erscheinungsdatum
2023
Seitenanfang
48
Seitenende
60
Publication
Elsevier BV
Erscheinungsdatum
2023
Zugänglichkeit
Rechte
Rechteangabe
© 2023 The Author(s)
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