RT Journal Article
T1 Enhanced Production of Herpes Simplex Virus 1 Amplicon Vectors by Gene Modification and Optimization of Packaging Cell Growth Medium
A1 Pérez Luz, Sara
A1 Fernández-Frías, Iván
A1 Díaz-Nido, Javier
AB 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.
PB Elsevier
YR 2020
FD 2020-03-10
LK https://hdl.handle.net/20.500.14352/102043
UL https://hdl.handle.net/20.500.14352/102043
LA eng
NO Author contributions: I.F.-F. performed the experiments. J.D.-N. conceived and designed the study. I.F.-F., S.P.-L., and J.D.-N. contributed to the interpretation of results. I.F.-F. and S.P.-L. wrote the first version of the paper. All the authors read and approved the final version of the manuscript.
DS Docta Complutense
RD 7 abr 2025