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Volume 21
Issue 17
10.3390/ijms21176011
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Article

Adenovirus-Mediated FasL Minigene Transfer Endows Transduced Cells with Killer Potential

by
Madalina Dumitrescu
1,
Violeta Georgeta Trusca
1,
Lorand Savu
2,
Ioana Georgeta Stancu
2,
Attila Cristian Ratiu
2,
Maya Simionescu
1 and
Anca Violeta Gafencu
1,*
1
Gene Regulation and Molecular Therapies Laboratory, Institute of Cellular Biology and Pathology “N. Simionescu”, 8, B.P. Hasdeu Street, 050568 Bucharest, Romania
2
Molecular Biology Department, Genetic Lab, 9 Căpitan Nicolae Drossu Street, 012071 Bucharest, Romania
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(17), 6011; https://doi.org/10.3390/ijms21176011
Submission received: 23 July 2020 / Revised: 17 August 2020 / Accepted: 18 August 2020 / Published: 20 August 2020
(This article belongs to the Section Molecular Biology)
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Abstract

Fas ligand (First apoptosis signal ligand, FasL, also known as CD95L) is the common executioner of apoptosis within the tumor necrosis factor (TNF) superfamily. We aimed to induce functional FasL expression in transduced cells using an adenovirus vector, which has the advantage of strong and transient induction of the gene included in the adenoviral genome. Here, we report that the adenovirus carrying a truncated FasL gene, named FasL minigene, encoding the full-length FasL protein (Ad-gFasL) is more efficient than the adenovirus carrying FasL cDNA (Ad-cFasL) in the induction of FasL expression in transduced cells. FasL minigene (2887 bp) lacking the second intron and a part of the 3′-UTR was created to reduce the gene length due to the size limitation of the adenoviral genome. The results show that, in transduced hepatocytes, strong expression of mRNA FasL appeared after 10 h for Ad-gFasL, while for Ad-cFasL, a faint expression appeared after 16 h. For Ad-gFasL, the protein expression was noticed starting with 0.5 transfection units (TU)/cell, while for Ad-cFasL, it could not be revealed. FasL-expressing endothelial cells induced apoptosis of A20 cells in co-culture experiments. FasL-expressing cells may be exploitable in various autoimmune diseases such as graft-versus-host disease, chronic colitis, and type I diabetes.
Keywords: FasL; minigene; cDNA; adenovirus; apoptosis FasL; minigene; cDNA; adenovirus; apoptosis
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MDPI and ACS Style

Dumitrescu, M.; Trusca, V.G.; Savu, L.; Stancu, I.G.; Ratiu, A.C.; Simionescu, M.; Gafencu, A.V. Adenovirus-Mediated FasL Minigene Transfer Endows Transduced Cells with Killer Potential. Int. J. Mol. Sci. 2020, 21, 6011. https://doi.org/10.3390/ijms21176011

AMA Style

Dumitrescu M, Trusca VG, Savu L, Stancu IG, Ratiu AC, Simionescu M, Gafencu AV. Adenovirus-Mediated FasL Minigene Transfer Endows Transduced Cells with Killer Potential. International Journal of Molecular Sciences. 2020; 21(17):6011. https://doi.org/10.3390/ijms21176011

Chicago/Turabian Style

Dumitrescu, Madalina, Violeta Georgeta Trusca, Lorand Savu, Ioana Georgeta Stancu, Attila Cristian Ratiu, Maya Simionescu, and Anca Violeta Gafencu. 2020. "Adenovirus-Mediated FasL Minigene Transfer Endows Transduced Cells with Killer Potential" International Journal of Molecular Sciences 21, no. 17: 6011. https://doi.org/10.3390/ijms21176011

APA Style

Dumitrescu, M., Trusca, V. G., Savu, L., Stancu, I. G., Ratiu, A. C., Simionescu, M., & Gafencu, A. V. (2020). Adenovirus-Mediated FasL Minigene Transfer Endows Transduced Cells with Killer Potential. International Journal of Molecular Sciences, 21(17), 6011. https://doi.org/10.3390/ijms21176011

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