Life Sciences & Biotechnology
Title : | Development of a novel dual luciferase reporter based high throughput assay for discovery of autophagy modulators and their functional characterization in mammalian system |
Area of research : | Life Sciences & Biotechnology |
Principal Investigator : | Dr. Jayanta Sarkar, CSIR-Central Drug Research Institute (CSIR-CDRI), Uttar Pradesh |
Timeline Start Year : | 2023 |
Timeline End Year : | 2026 |
Contact info : | j_sarkar@cdri.res.in |
Equipments : | Refrigerator ( above 950L volume)
Western blotting system with power pack |
Details
Executive Summary : | Autophagy is an evolutionarily conserved pathway involved in cellular homeostasis through lysosome mediated degradation of intracellular aggregation prone proteins, damaged organelles and various other unwanted macromolecules. Dysregulation of autophagy has been shown to be associated with various pathological conditions including cancer, metabolic syndromes, infectious diseases, neurodegenerative disorders and many other ailments. Therefore, modulation of autophagy has been emerged as a promising strategy for treatment of above disease conditions. There are numerous autophagy modulators which are at various stages of clinical development for treatment of several human ailments. Several biological assays are available for identification of autophagy modulators for therapeutic implications. These assays are based on wide variety of read out detection techniques including fluorescence/high content microscopy, flow cytometry, immunoblotting, microplate reader (for fluorescence/luminescence) etc. However, most of these assays suffer from limitations of either lower sensitivity or little scope for scaling up into high throughput format. Keeping this in mind, here we propose to develop a recombinant luciferase based enzymatic probes with high throughput applications for screening of autophagy modulators by determining autophagic flux. The assay is anticipated to be more sensitive, relatively easy to perform and can be carried out by using conventional laboratory equipment. The assay is also aimed at validating autophagy modulatory effect of identified molecule both in vitro and in vivo. Finally, potential hit molecules will be subjected to functional characterization in order to explore molecular mechanism of its autophagy modulatory effect. |
Total Budget (INR): | 47,67,724 |
Organizations involved