A thermogenetic tool enabling thermodynamic control of cellular protein functions

2023 Fiscal Year Final Research Report

• Project/Area Number: 22K20529
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0501:Physical chemistry, functional solid state chemistry, organic chemistry, polymers, organic materials, biomolecular chemistry, and related fields
• Research Institution: Kanazawa University
• Principal Investigator: VU QUANG CONG 金沢大学, ナノ生命科学研究所, 特任助教 (50961629)
• Project Period (FY): 2022-08-31 – 2024-03-31
• Keywords: Thermogenetics / Caspase 8

Outline of Final Research Achievements

This study aimed to develop a thermogenetic tool that utilizes heat to control protein functions for the manipulation of cellular processes. For the proof of concept, we developed a heat-activated human caspase 8 for programmed cell death. We fused a temperature-sensing domain with a human caspase 8 and expressed it in HEK293T cells. Upon heating, the temperature-sensing domain underwent coacervation, bringing two caspase 8 molecules into close proximity to induce dimerization and activation, thereby promoting cell death. For control, we introduced a C360A mutation in caspase 8 to inhibit its activation. When heated, HEK293T cells expressing the caspase 8 mutant remained alive, and cell division occurred. These results indicate the successful development of a heat-activated human caspase 8 for programmed cell death. Additionally, we developed a caspase 8 indicator to monitor its activation.

Free Research Field

Biotechnology

Academic Significance and Societal Importance of the Research Achievements

Our heat-activated caspase 8 system offers a precise and controllable method to initiate apoptosis by heating, with potential applications in targeted cancer therapies. The successful implementation of this approach validates thermogenetics and opens avenues for novel therapeutic strategies.