Luciferase-based reporting of suicide gene activity in murine mesenchymal stem cells

Gerace, D; Martiniello-Wilks, R; Habib, R; Simpson, M

Luciferase-based reporting of suicide gene activity in murine mesenchymal stem cells

Gerace, D

Martiniello-Wilks, R

Habib, R Simpson, M

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Publication Type:: Journal Article
Citation:: PLoS ONE, 2019, 14 (7)
Issue Date:: 2019-01-01

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Field	Value	Language
dc.contributor.author	Gerace, D https://orcid.org/0000-0001-9154-0624	en_US
dc.contributor.author	Martiniello-Wilks, R https://orcid.org/0000-0002-0038-3430	en_US
dc.contributor.author	Habib, R	en_US
dc.contributor.author	Simpson, M	en_US
dc.date.available	2019-07-05	en_US
dc.date.issued	2019-01-01	en_US
dc.identifier.citation	PLoS ONE, 2019, 14 (7)	en_US
dc.identifier.uri	http://hdl.handle.net/10453/135900
dc.description.abstract	© 2019 Gerace et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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.	en_US
dc.relation.ispartof	PLoS ONE	en_US
dc.relation.isbasedon	10.1371/journal.pone.0220013	en_US
dc.subject.classification	General Science & Technology	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Biomarkers	en_US
dc.subject.mesh	Cell Line	en_US
dc.subject.mesh	Gene Expression	en_US
dc.subject.mesh	Genes, Reporter	en_US
dc.subject.mesh	Genes, Transgenic, Suicide	en_US
dc.subject.mesh	Immunophenotyping	en_US
dc.subject.mesh	Luciferases	en_US
dc.subject.mesh	Mesenchymal Stem Cells	en_US
dc.subject.mesh	Mice	en_US
dc.title	Luciferase-based reporting of suicide gene activity in murine mesenchymal stem cells	en_US
dc.type	Journal Article
utslib.citation.volume	7	en_US
utslib.citation.volume	14	en_US
utslib.for	0604 Genetics	en_US
utslib.for	0601 Biochemistry and Cell Biology	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney/Strength - CHT - Health Technologies
pubs.organisational-group	/University of Technology Sydney/Students
utslib.copyright.status	open_access
pubs.issue	7	en_US
pubs.publication-status	Published	en_US
pubs.volume	14	en_US

Abstract:

© 2019 Gerace et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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.

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http://hdl.handle.net/10453/135900