| 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
|
| Project Status |
Completed (Fiscal Year 2023)
|
| Budget Amount *help |
¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2023: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
|
|---|
| Keywords | Thermogenetics / Caspase 8 / Biosensors / Bioimaging / Fluorescence thermometry / FLIM |
|---|
| Outline of Research at the Start |
Thermogenetics employ thermosensitive proteins to manipulate protein activities by changing temperature. Here, I aim to create a thermogenetic tool to thermodynamically control the activity of protein inside live cells by employing the phase transition behaviors of thermal-responsive polypeptides
|
| 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.
|
| 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.
|
