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Luciferase-based reporting of suicide gene activity in murine mesenchymal stem cells.
PLoS One 2019; 14(7):e0220013Plos

Abstract

Due to their ease of isolation, gene modification and tumor-homing properties, mesenchymal stem cells (MSCs) are an attractive cellular vehicle for the delivery of toxic suicide genes to a variety of cancers in pre-clinical models. In addition, the incorporation of suicide genes in stem cell-derived cell replacement therapies improves their safety profile by permitting graft destruction in the event of unexpected tumorigeneses or unwanted differentiation. Due to the functional requirement of ATP for the Firefly luciferase gene Luc2 to produce light, luciferase-based reporting of cytotoxicity can be engineered into potential cell therapies. Consequently, we nucleofected mammalian expression plasmids containing both the Luc2 and the yeast fusion cytosine deaminase uracil phosphoribosyltransferase (CDUPRT) genes for expression in murine MSCs to assess luciferase as a reporter of suicide gene cytotoxicity, and MSC as vehicles of suicide gene therapy. In vitro bioluminescence imaging (BLI) showed that following the addition of the non-toxic prodrug fluorocytosine (5-FC), CDUPRT-expressing MSCs displayed enhanced cytotoxicity in comparison to Luc2 reporter MSC controls. This study demonstrates the utility of luciferase as a reporter of CDUPRT-mediated cytotoxicity in murine MSC using BLI.

Authors+Show Affiliations

The School of Life Sciences and the Centre for Health Technologies, University of Technology Sydney, Sydney, Australia.The School of Life Sciences and the Centre for Health Technologies, University of Technology Sydney, Sydney, Australia. Translational Cancer Research Group, University of Technology Sydney, Sydney, Australia.The School of Life Sciences and the Centre for Health Technologies, University of Technology Sydney, Sydney, Australia.The School of Life Sciences and the Centre for Health Technologies, University of Technology Sydney, Sydney, Australia.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31318955

Citation

Gerace, Dario, et al. "Luciferase-based Reporting of Suicide Gene Activity in Murine Mesenchymal Stem Cells." PloS One, vol. 14, no. 7, 2019, pp. e0220013.
Gerace D, Martiniello-Wilks R, Habib R, et al. Luciferase-based reporting of suicide gene activity in murine mesenchymal stem cells. PLoS ONE. 2019;14(7):e0220013.
Gerace, D., Martiniello-Wilks, R., Habib, R., & Simpson, A. M. (2019). Luciferase-based reporting of suicide gene activity in murine mesenchymal stem cells. PloS One, 14(7), pp. e0220013. doi:10.1371/journal.pone.0220013.
Gerace D, et al. Luciferase-based Reporting of Suicide Gene Activity in Murine Mesenchymal Stem Cells. PLoS ONE. 2019;14(7):e0220013. PubMed PMID: 31318955.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Luciferase-based reporting of suicide gene activity in murine mesenchymal stem cells. AU - Gerace,Dario, AU - Martiniello-Wilks,Rosetta, AU - Habib,Rosaline, AU - Simpson,Ann Margaret, Y1 - 2019/07/18/ PY - 2019/05/14/received PY - 2019/07/05/accepted PY - 2019/7/19/entrez PY - 2019/7/19/pubmed PY - 2019/7/19/medline SP - e0220013 EP - e0220013 JF - PloS one JO - PLoS ONE VL - 14 IS - 7 N2 - Due to their ease of isolation, gene modification and tumor-homing properties, mesenchymal stem cells (MSCs) are an attractive cellular vehicle for the delivery of toxic suicide genes to a variety of cancers in pre-clinical models. In addition, the incorporation of suicide genes in stem cell-derived cell replacement therapies improves their safety profile by permitting graft destruction in the event of unexpected tumorigeneses or unwanted differentiation. Due to the functional requirement of ATP for the Firefly luciferase gene Luc2 to produce light, luciferase-based reporting of cytotoxicity can be engineered into potential cell therapies. Consequently, we nucleofected mammalian expression plasmids containing both the Luc2 and the yeast fusion cytosine deaminase uracil phosphoribosyltransferase (CDUPRT) genes for expression in murine MSCs to assess luciferase as a reporter of suicide gene cytotoxicity, and MSC as vehicles of suicide gene therapy. In vitro bioluminescence imaging (BLI) showed that following the addition of the non-toxic prodrug fluorocytosine (5-FC), CDUPRT-expressing MSCs displayed enhanced cytotoxicity in comparison to Luc2 reporter MSC controls. This study demonstrates the utility of luciferase as a reporter of CDUPRT-mediated cytotoxicity in murine MSC using BLI. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/31318955/Luciferase-based_reporting_of_suicide_gene_activity_in_murine_mesenchymal_stem_cells L2 - http://dx.plos.org/10.1371/journal.pone.0220013 DB - PRIME DP - Unbound Medicine ER -