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Developing a circularly permuted variant of Renilla luciferase as a bioluminescent sensor for measuring Caspase-9 activity in the cell-free and cell-based systems

Mokhtar-Ahmadabadi, Roya and Madadi, Zahra and Akbari-Birgani, Shiva and Grillon, Catherine and Hasani, Leila and Hosseinkhani, Saman and Zareian, Shekufeh (2018) Developing a circularly permuted variant of Renilla luciferase as a bioluminescent sensor for measuring Caspase-9 activity in the cell-free and cell-based systems. Biochemical and Biophysical Research Communications, 506 (4). pp. 1032-1039. ISSN 0006-291X. http://resolver.caltech.edu/CaltechAUTHORS:20181106-100856476

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Abstract

Biosensors and whole cell biosensors consisting of biological molecules and living cells can sense a special stimulus on a living system and convert it to a measurable signal. A major group of them are the bioluminescent sensors derived from luciferases. This type of biosensors has a broad application in molecular biology and imaging systems. In this project, a luciferase-based biosensor for detecting and measuring caspase-9 activity is designed and constructed using the circular permutation strategy. The spectroscopic method results reveal changes in the biosensor structure. Additionally, its activity is examined in a cell-free coupled assay system. Afterward, the biosensor is utilized for measuring the cellular caspase-9 activity upon apoptosis induction in a cancer cell line. In following the gene of biosensor is sub-cloned into a eukaryotic vector and transfected to HEK293T cell line and then its activity is measured upon apoptosis induction in the presence and absence of a caspase-9 inhibitor. The obtained results show that the designed biosensor detects the caspase-9 activity in the cell-free and cell-based systems.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.bbrc.2018.11.009DOIArticle
ORCID:
AuthorORCID
Akbari-Birgani, Shiva0000-0001-8826-4083
Additional Information:© 2018 Elsevier Inc. Received 26 October 2018, Accepted 3 November 2018, Available online 6 November 2018. This work is supported by the Institute for Advanced Studies in Basic Sciences (IASBS). Financial support for this work is provided by Research Council of Institute for Advanced Studies in Basic Sciences (IASBS). We gratefully appreciate centre de biophysique moléculaire cnrs, France, for dedicating facilities, and the scientific collaboration to us.
Funders:
Funding AgencyGrant Number
Institute for Advanced Studies in Basic SciencesUNSPECIFIED
Record Number:CaltechAUTHORS:20181106-100856476
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20181106-100856476
Official Citation:Roya Mokhtar-Ahmadabadi, Zahra Madadi, Shiva Akbari-Birgani, Catherine Grillon, Leila Hasani, Saman Hosseinkhani, Shekufeh Zareian, Developing a circularly permuted variant of Renilla luciferase as a bioluminescent sensor for measuring Caspase-9 activity in the cell-free and cell-based systems, Biochemical and Biophysical Research Communications, Volume 506, Issue 4, 2018, Pages 1032-1039, ISSN 0006-291X, https://doi.org/10.1016/j.bbrc.2018.11.009. (http://www.sciencedirect.com/science/article/pii/S0006291X18324070)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:90665
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:06 Nov 2018 18:20
Last Modified:20 Nov 2018 18:37

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