GAMER-Ad: a novel and rapid method for generating recombinant adenoviruses
Oncolytic adenoviruses have become ideal agents in the path toward treating cancer. Such viruses have been engineered to conditionally replicate in malignant cells in which certain signaling pathways have been disrupted. Other than such oncolytic properties, the viruses need to activate the immune s...
Main Authors: | , , , , , , , , , , |
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Format: | Article |
Language: | English |
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Elsevier
2021-03-01
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Series: | Molecular Therapy: Methods & Clinical Development |
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Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050121000140 |
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doaj-7934eb39ade74b8590b02cd788167162 |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Firas Hamdan Beatriz Martins Michaela Feodoroff Yvonne Giannoula Sara Feola Manlio Fusciello Jacopo Chiaro Gabriella Antignani Mikaela Grönholm Erkko Ylösmäki Vincenzo Cerullo |
spellingShingle |
Firas Hamdan Beatriz Martins Michaela Feodoroff Yvonne Giannoula Sara Feola Manlio Fusciello Jacopo Chiaro Gabriella Antignani Mikaela Grönholm Erkko Ylösmäki Vincenzo Cerullo GAMER-Ad: a novel and rapid method for generating recombinant adenoviruses Molecular Therapy: Methods & Clinical Development adenoviruses Gibson Assembly gene therapy oncolytic viruses molecular cloning |
author_facet |
Firas Hamdan Beatriz Martins Michaela Feodoroff Yvonne Giannoula Sara Feola Manlio Fusciello Jacopo Chiaro Gabriella Antignani Mikaela Grönholm Erkko Ylösmäki Vincenzo Cerullo |
author_sort |
Firas Hamdan |
title |
GAMER-Ad: a novel and rapid method for generating recombinant adenoviruses |
title_short |
GAMER-Ad: a novel and rapid method for generating recombinant adenoviruses |
title_full |
GAMER-Ad: a novel and rapid method for generating recombinant adenoviruses |
title_fullStr |
GAMER-Ad: a novel and rapid method for generating recombinant adenoviruses |
title_full_unstemmed |
GAMER-Ad: a novel and rapid method for generating recombinant adenoviruses |
title_sort |
gamer-ad: a novel and rapid method for generating recombinant adenoviruses |
publisher |
Elsevier |
series |
Molecular Therapy: Methods & Clinical Development |
issn |
2329-0501 |
publishDate |
2021-03-01 |
description |
Oncolytic adenoviruses have become ideal agents in the path toward treating cancer. Such viruses have been engineered to conditionally replicate in malignant cells in which certain signaling pathways have been disrupted. Other than such oncolytic properties, the viruses need to activate the immune system in order to sustain a long-term response. Therefore, oncolytic adenoviruses have been genetically modified to express various immune-stimulatory agents to achieve this. However, genetically modifying adenoviruses is very time consuming and labor intensive with the current available methods. In this paper, we describe a novel method we have called GAMER-Ad to genetically modify adenovirus genomes within 2 days. Our method entails the replacement of the gp19k gene in the E3 region with any given gene of interest (GOI) using Gibson Assembly avoiding the homologous recombination between the shuttle and the parental plasmid. In this manuscript as proof of concept we constructed and characterized three oncolytic adenoviruses expressing CXCL9, CXCL10, and interleukin-15 (IL-15). We demonstrate that our novel method is fast, reliable, and simple compared to other methods. We anticipate that our method will be used in the future to genetically engineer oncolytic but also other adenoviruses used for gene therapy as well. |
topic |
adenoviruses Gibson Assembly gene therapy oncolytic viruses molecular cloning |
url |
http://www.sciencedirect.com/science/article/pii/S2329050121000140 |
work_keys_str_mv |
AT firashamdan gameradanovelandrapidmethodforgeneratingrecombinantadenoviruses AT beatrizmartins gameradanovelandrapidmethodforgeneratingrecombinantadenoviruses AT michaelafeodoroff gameradanovelandrapidmethodforgeneratingrecombinantadenoviruses AT yvonnegiannoula gameradanovelandrapidmethodforgeneratingrecombinantadenoviruses AT sarafeola gameradanovelandrapidmethodforgeneratingrecombinantadenoviruses AT manliofusciello gameradanovelandrapidmethodforgeneratingrecombinantadenoviruses AT jacopochiaro gameradanovelandrapidmethodforgeneratingrecombinantadenoviruses AT gabriellaantignani gameradanovelandrapidmethodforgeneratingrecombinantadenoviruses AT mikaelagronholm gameradanovelandrapidmethodforgeneratingrecombinantadenoviruses AT erkkoylosmaki gameradanovelandrapidmethodforgeneratingrecombinantadenoviruses AT vincenzocerullo gameradanovelandrapidmethodforgeneratingrecombinantadenoviruses |
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1724222177752907776 |
spelling |
doaj-7934eb39ade74b8590b02cd7881671622021-03-13T04:23:53ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012021-03-0120625634GAMER-Ad: a novel and rapid method for generating recombinant adenovirusesFiras Hamdan0Beatriz Martins1Michaela Feodoroff2Yvonne Giannoula3Sara Feola4Manlio Fusciello5Jacopo Chiaro6Gabriella Antignani7Mikaela Grönholm8Erkko Ylösmäki9Vincenzo Cerullo10Laboratory of Immunovirotherapy (IVTLab), Drug Research Program, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5E, 00790 Helsinki, Finland; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland; TRIMM, Translational Immunology Research Program, University of Helsinki, Helsinki, FinlandLaboratory of Immunovirotherapy (IVTLab), Drug Research Program, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5E, 00790 Helsinki, Finland; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland; TRIMM, Translational Immunology Research Program, University of Helsinki, Helsinki, FinlandLaboratory of Immunovirotherapy (IVTLab), Drug Research Program, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5E, 00790 Helsinki, Finland; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland; TRIMM, Translational Immunology Research Program, University of Helsinki, Helsinki, FinlandLaboratory of Immunovirotherapy (IVTLab), Drug Research Program, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5E, 00790 Helsinki, Finland; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland; TRIMM, Translational Immunology Research Program, University of Helsinki, Helsinki, FinlandLaboratory of Immunovirotherapy (IVTLab), Drug Research Program, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5E, 00790 Helsinki, Finland; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland; TRIMM, Translational Immunology Research Program, University of Helsinki, Helsinki, FinlandLaboratory of Immunovirotherapy (IVTLab), Drug Research Program, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5E, 00790 Helsinki, Finland; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland; TRIMM, Translational Immunology Research Program, University of Helsinki, Helsinki, FinlandLaboratory of Immunovirotherapy (IVTLab), Drug Research Program, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5E, 00790 Helsinki, Finland; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland; TRIMM, Translational Immunology Research Program, University of Helsinki, Helsinki, FinlandLaboratory of Immunovirotherapy (IVTLab), Drug Research Program, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5E, 00790 Helsinki, Finland; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland; TRIMM, Translational Immunology Research Program, University of Helsinki, Helsinki, FinlandLaboratory of Immunovirotherapy (IVTLab), Drug Research Program, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5E, 00790 Helsinki, Finland; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland; TRIMM, Translational Immunology Research Program, University of Helsinki, Helsinki, Finland; iCAN Digital Precision Cancer Medicine Flagship, University of Helsinki, Helsinki, FinlandLaboratory of Immunovirotherapy (IVTLab), Drug Research Program, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5E, 00790 Helsinki, Finland; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland; TRIMM, Translational Immunology Research Program, University of Helsinki, Helsinki, FinlandLaboratory of Immunovirotherapy (IVTLab), Drug Research Program, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5E, 00790 Helsinki, Finland; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland; TRIMM, Translational Immunology Research Program, University of Helsinki, Helsinki, Finland; iCAN Digital Precision Cancer Medicine Flagship, University of Helsinki, Helsinki, Finland; Department of Molecular Medicine and Medical Biotechnology and CEINGE, Naples University 24 Federico II, 80131 Naples, Italy; Corresponding author: Vincenzo Cerullo, Laboratory of Immunovirotherapy (IVTLab), Drug Research Program, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5E, 00790 Helsinki, Finland.Oncolytic adenoviruses have become ideal agents in the path toward treating cancer. Such viruses have been engineered to conditionally replicate in malignant cells in which certain signaling pathways have been disrupted. Other than such oncolytic properties, the viruses need to activate the immune system in order to sustain a long-term response. Therefore, oncolytic adenoviruses have been genetically modified to express various immune-stimulatory agents to achieve this. However, genetically modifying adenoviruses is very time consuming and labor intensive with the current available methods. In this paper, we describe a novel method we have called GAMER-Ad to genetically modify adenovirus genomes within 2 days. Our method entails the replacement of the gp19k gene in the E3 region with any given gene of interest (GOI) using Gibson Assembly avoiding the homologous recombination between the shuttle and the parental plasmid. In this manuscript as proof of concept we constructed and characterized three oncolytic adenoviruses expressing CXCL9, CXCL10, and interleukin-15 (IL-15). We demonstrate that our novel method is fast, reliable, and simple compared to other methods. We anticipate that our method will be used in the future to genetically engineer oncolytic but also other adenoviruses used for gene therapy as well.http://www.sciencedirect.com/science/article/pii/S2329050121000140adenovirusesGibson Assemblygene therapyoncolytic virusesmolecular cloning |