A novel thermostable beetle luciferase based cytotoxicity assay

Abstract Cytotoxicity assays are essential for the testing and development of novel immunotherapies for the treatment of cancer. We recently described a novel cytotoxicity assay, termed the Matador assay, which was based on marine luciferases and their engineered derivatives. In this study, we descr...

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Autores principales:	Sunju Choi, Hittu Matta, Ramakrishnan Gopalakrishnan, Venkatesh Natarajan, Songjie Gong, Alberto Jeronimo, Wei-Ying Kuo, Bryant Bravo, Preet M. Chaudhary
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/bc13b55b93244e408c8f65821ee6c580
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spelling	oai:doaj.org-article:bc13b55b93244e408c8f65821ee6c5802021-12-02T14:35:47ZA novel thermostable beetle luciferase based cytotoxicity assay10.1038/s41598-021-89404-z2045-2322https://doaj.org/article/bc13b55b93244e408c8f65821ee6c5802021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89404-zhttps://doaj.org/toc/2045-2322Abstract Cytotoxicity assays are essential for the testing and development of novel immunotherapies for the treatment of cancer. We recently described a novel cytotoxicity assay, termed the Matador assay, which was based on marine luciferases and their engineered derivatives. In this study, we describe the development of a new cytotoxicity assay termed ‘Matador-Glo assay’ which takes advantage of a thermostable variant of Click Beetle Luciferase (Luc146-1H2). Matador-Glo assay utilizes Luc146-1H2 and D-luciferin as the luciferase-substrate pair for luminescence detection. The assay involves ectopic over-expression of Luc146-1H2 in the cytosol of target cells of interest. Upon damage to the membrane integrity, the Luc146-1H2 is either released from the dead and dying cells or its activity is preferentially measured in dead and dying cells. We demonstrate that this assay is simple, fast, specific, sensitive, cost-efficient, and not labor-intensive. We further demonstrate that the Matador-Glo assay can be combined with the marine luciferase-based Matador assay to develop a dual luciferase assay for cell death detection. Finally, we demonstrate that the Luc146-1H2 expressing target cells can also be used for in vivo bioluminescence imaging applications.Sunju ChoiHittu MattaRamakrishnan GopalakrishnanVenkatesh NatarajanSongjie GongAlberto JeronimoWei-Ying KuoBryant BravoPreet M. ChaudharyNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sunju Choi Hittu Matta Ramakrishnan Gopalakrishnan Venkatesh Natarajan Songjie Gong Alberto Jeronimo Wei-Ying Kuo Bryant Bravo Preet M. Chaudhary A novel thermostable beetle luciferase based cytotoxicity assay
description	Abstract Cytotoxicity assays are essential for the testing and development of novel immunotherapies for the treatment of cancer. We recently described a novel cytotoxicity assay, termed the Matador assay, which was based on marine luciferases and their engineered derivatives. In this study, we describe the development of a new cytotoxicity assay termed ‘Matador-Glo assay’ which takes advantage of a thermostable variant of Click Beetle Luciferase (Luc146-1H2). Matador-Glo assay utilizes Luc146-1H2 and D-luciferin as the luciferase-substrate pair for luminescence detection. The assay involves ectopic over-expression of Luc146-1H2 in the cytosol of target cells of interest. Upon damage to the membrane integrity, the Luc146-1H2 is either released from the dead and dying cells or its activity is preferentially measured in dead and dying cells. We demonstrate that this assay is simple, fast, specific, sensitive, cost-efficient, and not labor-intensive. We further demonstrate that the Matador-Glo assay can be combined with the marine luciferase-based Matador assay to develop a dual luciferase assay for cell death detection. Finally, we demonstrate that the Luc146-1H2 expressing target cells can also be used for in vivo bioluminescence imaging applications.
format	article
author	Sunju Choi Hittu Matta Ramakrishnan Gopalakrishnan Venkatesh Natarajan Songjie Gong Alberto Jeronimo Wei-Ying Kuo Bryant Bravo Preet M. Chaudhary
author_facet	Sunju Choi Hittu Matta Ramakrishnan Gopalakrishnan Venkatesh Natarajan Songjie Gong Alberto Jeronimo Wei-Ying Kuo Bryant Bravo Preet M. Chaudhary
author_sort	Sunju Choi
title	A novel thermostable beetle luciferase based cytotoxicity assay
title_short	A novel thermostable beetle luciferase based cytotoxicity assay
title_full	A novel thermostable beetle luciferase based cytotoxicity assay
title_fullStr	A novel thermostable beetle luciferase based cytotoxicity assay
title_full_unstemmed	A novel thermostable beetle luciferase based cytotoxicity assay
title_sort	novel thermostable beetle luciferase based cytotoxicity assay
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/bc13b55b93244e408c8f65821ee6c580
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A novel thermostable beetle luciferase based cytotoxicity assay

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