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Viruses 2018, 10(1), 50; doi:10.3390/v10010050

CRISPR/Cas9—Advancing Orthopoxvirus Genome Editing for Vaccine and Vector Development

Biosafety of Genome Editing Research Group, GenØk-Centre for Biosafety, Siva Innovation Centre, N-9294 Tromsø, Norway
Artic Infection Biology, Department of Artic and Marine Biology, The Artic University of Norway, N-9037 Tromsø, Norway
Molecular Inflammation Research Group, Institute of Medical Biology, The Arctic University of Norway, N-9037 Tromsø, Norway
Author to whom correspondence should be addressed.
Received: 13 December 2017 / Revised: 17 January 2018 / Accepted: 21 January 2018 / Published: 22 January 2018
(This article belongs to the Special Issue Applications of CRISPR Technology in Virology 2018)
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The clustered regularly interspaced short palindromic repeat (CRISPR)/associated protein 9 (Cas9) technology is revolutionizing genome editing approaches. Its high efficiency, specificity, versatility, flexibility, simplicity and low cost have made the CRISPR/Cas9 system preferable to other guided site-specific nuclease-based systems such as TALENs (Transcription Activator-like Effector Nucleases) and ZFNs (Zinc Finger Nucleases) in genome editing of viruses. CRISPR/Cas9 is presently being applied in constructing viral mutants, preventing virus infections, eradicating proviral DNA, and inhibiting viral replication in infected cells. The successful adaptation of CRISPR/Cas9 to editing the genome of Vaccinia virus paves the way for its application in editing other vaccine/vector-relevant orthopoxvirus (OPXV) strains. Thus, CRISPR/Cas9 can be used to resolve some of the major hindrances to the development of OPXV-based recombinant vaccines and vectors, including sub-optimal immunogenicity; transgene and genome instability; reversion of attenuation; potential of spread of transgenes to wildtype strains and close contacts, which are important biosafety and risk assessment considerations. In this article, we review the published literature on the application of CRISPR/Cas9 in virus genome editing and discuss the potentials of CRISPR/Cas9 in advancing OPXV-based recombinant vaccines and vectors. We also discuss the application of CRISPR/Cas9 in combating viruses of clinical relevance, the limitations of CRISPR/Cas9 and the current strategies to overcome them. View Full-Text
Keywords: CRISPR/Cas9; genome editing; modified Vaccinia virus Ankara; orthopoxvirus; recombination; risk assessment; site-specific; Vaccinia virus; vaccine; vector CRISPR/Cas9; genome editing; modified Vaccinia virus Ankara; orthopoxvirus; recombination; risk assessment; site-specific; Vaccinia virus; vaccine; vector

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Okoli, A.; Okeke, M.I.; Tryland, M.; Moens, U. CRISPR/Cas9—Advancing Orthopoxvirus Genome Editing for Vaccine and Vector Development. Viruses 2018, 10, 50.

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