Enhanced production of herpes simplex virus 1 amplicon vectors by gene modification and optimization of packaging cell growth medium
Entity
UAM. Departamento de Biología MolecularPublisher
Cell PressDate
2020-06-12Citation
10.1016/j.omtm.2020.03.005
Molecular Therapy: Methods & Clinical Development 17 (2020): 491-496
ISSN
2329-0501 (online)DOI
10.1016/j.omtm.2020.03.005Funded by
This work was supported by grants of the Spanish National Research Plan (SAF 2015–69361-R), Friedreich Ataxia Research Alliance (FARA), FARA Ireland, Spanish FEDAES, GENEFA, and Babel FamilyProject
Gobierno de España. SAF 2015–69361-REditor's Version
https://doi.org/10.1016/j.omtm.2020.03.005Subjects
Gene therapy; HSV-1 amplicons; Bacterial artificial chromosomes; BACs; Bcl-2; Apoptosis; Media supplementation; Biología y Biomedicina / BiologíaRights
© 2020 The AuthorsEsta obra está bajo una licencia de Creative Commons Reconocimiento-NoComercial-SinObraDerivada 4.0 Internacional.
Abstract
Herpes simplex virus 1 (HSV-1)-derived amplicon vectors are
unique in their ability to accommodate large DNA molecules
allowing whole genomic loci to be included with all of their regulatory
elements. Additional advantages of these amplicons
include their minimal toxicity and ability to persist as episomes,
with negligible risk of insertional mutagenesis, being
particularly well-suited for gene therapy of neurological disorders
due to their outstanding ability to deliver genes into neurons
and other neural cells. However, extensive gene therapy
application has been hindered by difficulties in vector production.
This work improved HSV-1 amplicons production by genetic
modification of the packaging cell line and optimization
of the culture medium. A stably-transfected Vero 2-2 cell line
overexpressing the anti-apoptotic Bcl-2 protein was generated,
exhibiting an increased resistance to apoptosis, prolonged culture
duration, and a significant improvement in viral vector
production. Additionally, supplementation of the growth medium
with antioxidants, polyamines, amino acids, and reduced
glutathione further increased the yield of packaged amplicon
vectors. With these modifications, HSV-1 amplicons could be
isolated from culture supernatants instead of cell lysates, leading
to vector preparations with higher titer and purity and
paving the way for generation of stable cell lines that are
capable of continuous herpesviral vector production
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Google Scholar:Fernández-Frías, Iván
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Pérez-Luz, Sara
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Díaz Nido, Javier
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