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      Use of Reporter Genes in the Generation of Vaccinia Virus-Derived Vectors

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          Abstract

          Vaccinia virus (VACV) is one of the most extensively-studied viruses of the Poxviridae family. It is easy to genetically modify, so it has become a key tool for many applications. In this context, reporter genes facilitate the study of the role of foreign genes introduced into the genome of VACV. In this review, we describe the type of reporter genes that have been used to generate reporter-expressing VACV and the applications of the recombinant viruses obtained. Reporter-expressing VACV are currently employed in basic and immunology research, in the development of vaccines and cancer treatment.

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          Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells.

          C M Gorman, L Moffat, B H HOWARD (1982)
          We constructed a series of recombinant genomes which directed expression of the enzyme chloramphenicol acetyltransferase (CAT) in mammalian cells. The prototype recombinant in this series, pSV2-cat, consisted of the beta-lactamase gene and origin of replication from pBR322 coupled to a simian virus 40 (SV40) early transcription region into which CAT coding sequences were inserted. Readily measured levels of CAT accumulated within 48 h after the introduction of pSV2-cat DNA into African green monkey kidney CV-1 cells. Because endogenous CAT activity is not present in CV-1 or other mammalian cells, and because rapid, sensitive assays for CAT activity are available, these recombinants provided a uniquely convenient system for monitoring the expression of foreign DNAs in tissue culture cells. To demonstrate the usefulness of this system, we constructed derivatives of pSV2-cat from which part or all of the SV40 promoter region was removed. Deletion of one copy of the 72-base-pair repeat sequence in the SV40 promoter caused no significant decrease in CAT synthesis in monkey kidney CV-1 cells; however, an additional deletion of 50 base pairs from the second copy of the repeats reduced CAT synthesis to 11% of its level in the wild type. We also constructed a recombinant, pSV0-cat, in which the entire SV40 promoter region was removed and a unique HindIII site was substituted for the insertion of other promoter sequences.
          Article 0 comments Cited 216 times – based on 0 reviews     Review now
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            Mammalian cell transfection: the present and the future

            Tae-Kyung Kim, James Eberwine (2010)
            Transfection is a powerful analytical tool enabling study of the function of genes and gene products in cells. The transfection methods are broadly classified into three groups; biological, chemical, and physical. These methods have advanced to make it possible to deliver nucleic acids to specific subcellular regions of cells by use of a precisely controlled laser-microcope system. The combination of point-directed transfection and mRNA transfection is a new way of studying the function of genes and gene products. However, each method has its own advantages and disadvantages so the optimum method depends on experimental design and objective.
            Article 0 comments Cited 204 times – based on 0 reviews     Review now
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              Vaccinia virus immune evasion: mechanisms, virulence and immunogenicity.

              Jeffrey Smith, Camilla Benfield, Stephen Ferguson … (2013)
              Virus infection of mammalian cells is sensed by pattern recognition receptors and leads to an innate immune response that restricts virus replication and induces adaptive immunity. In response, viruses have evolved many countermeasures that enable them to replicate and be transmitted to new hosts, despite the host innate immune response. Poxviruses, such as vaccinia virus (VACV), have large DNA genomes and encode many proteins that are dedicated to host immune evasion. Some of these proteins are secreted from the infected cell, where they bind and neutralize complement factors, interferons, cytokines and chemokines. Other VACV proteins function inside cells to inhibit apoptosis or signalling pathways that lead to the production of interferons and pro-inflammatory cytokines and chemokines. In this review, these VACV immunomodulatory proteins are described and the potential to create more immunogenic VACV strains by manipulation of the gene encoding these proteins is discussed.
              Article 0 comments Cited 160 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                Luis Martinez-Sobrido: Role: Academic Editor
                Journal
                Journal ID (nlm-ta): Viruses
                Journal ID (iso-abbrev): Viruses
                Journal ID (publisher-id): viruses
                Title: Viruses
                Publisher: MDPI
                ISSN (Electronic): 1999-4915
                Publication date (Electronic): 21 May 2016
                Publication date Collection: May 2016
                Volume: 8
                Issue: 5
                Electronic Location Identifier: 134
                Affiliations
                Department of Preventive Medicine, Public Health and Microbiology, Universidad Autónoma, E-28029 Madrid, Spain; sally.alali@ 123456yahoo.com (S.A.A.); sara.baldanta@ 123456uam.es (S.B.); mercedes.fernandeze@ 123456uam.es (M.F.-E.)
                Author notes
                [* ]Correspondence: susana.guerra@ 123456uam.es ; Tel.: +34-91-497-5440
                [†]

                These authors contributed equally to this work.

                Article
                Publisher ID: viruses-08-00134
                DOI: 10.3390/v8050134
                PMC ID: 4885089
                PubMed ID: 27213433
                SO-VID: 34cc6d90-d09c-487b-962b-10c4bfdc748e
                Copyright © © 2016 by the authors; licensee MDPI, Basel, Switzerland.
                License:

                This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                Date received : 18 March 2016
                Date accepted : 12 May 2016
                Categories
                Subject: Review

                ScienceOpen disciplines: Microbiology & Virology
                Keywords: vaccinia virus,vaccine,reporter-expressing virus
                Data availability:
                ScienceOpen disciplines: Microbiology & Virology
                Keywords: vaccinia virus, vaccine, reporter-expressing virus

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