Development of Novel Bioluminescent Biosensors Monitoring the Conformation and Activity of the Merlin Tumour Suppressor
Abstract
:1. Introduction
2. Results
2.1. Development and Validation of Mer-Intra-BS
2.2. Live Cell Bioluminescent Imaging (BLI) Analysis of Mer-Intra-BS Activity
2.3. Cell Density Dependent Regulation of Mer-Intra-BS Activity
2.4. Characterization of Mer-Intra-BS In Vitro
3. Discussion
4. Materials and Methods
4.1. Biosensor Design and Construction
4.2. Site-Directed Mutagenesis
4.3. Cell Culture
4.4. Protein Extraction and Western Blot Analysis
4.5. Lentivirus Production and Stable Cell Line Generation
4.6. Luciferase Assays
4.7. Bioluminescent Imaging (BLI) Analysis
4.8. Monitoring Cell Density Dependent Activation of Mer-Intra-BS
4.9. Purification of His-Tagged Mer-Intra-BS
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Pipchuk, A.; Kelly, T.; Carew, M.; Nicol, C.; Yang, X. Development of Novel Bioluminescent Biosensors Monitoring the Conformation and Activity of the Merlin Tumour Suppressor. Int. J. Mol. Sci. 2024, 25, 1527. https://doi.org/10.3390/ijms25031527
Pipchuk A, Kelly T, Carew M, Nicol C, Yang X. Development of Novel Bioluminescent Biosensors Monitoring the Conformation and Activity of the Merlin Tumour Suppressor. International Journal of Molecular Sciences. 2024; 25(3):1527. https://doi.org/10.3390/ijms25031527
Chicago/Turabian StylePipchuk, Alexander, Tynan Kelly, Madeleine Carew, Christopher Nicol, and Xiaolong Yang. 2024. "Development of Novel Bioluminescent Biosensors Monitoring the Conformation and Activity of the Merlin Tumour Suppressor" International Journal of Molecular Sciences 25, no. 3: 1527. https://doi.org/10.3390/ijms25031527