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Thermogenetics enables precise cellular control using heat-activated proteins. Researchers developed a heat-inducible programmed cell death system using temperature-responsive elastin-like polypeptides (ELPs) fused to caspase 8 (CASP8).

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Cell Biology

Background:

  • Thermogenetics offers advantages over optogenetics for cellular control due to heat's superior penetration.
  • Developing targeted methods for inducing cellular responses like programmed cell death is crucial for biological research and therapeutics.

Purpose of the Study:

  • To engineer a thermogenetically controlled system for activating human caspase 8 (CASP8) and inducing programmed cell death.
  • To utilize temperature-responsive elastin-like polypeptides (ELPs) for heat-activated protein function.

Main Methods:

  • Constructed a fusion protein combining ELP[V60] with the catalytic domain of CASP8 (ELP[V60]-CASP8).
  • Exploited the reversible phase transition of ELPs for localized protein coacervation and activation upon heating.
  • Utilized optical heating with a 1470 nm laser and fluorescence lifetime-based thermometry for precise, localized CASP8 activation in single cells.

Main Results:

  • The ELP[V60]-CASP8 fusion protein induced cell death in HEK293T cells at temperatures above 35 °C.
  • Higher temperatures correlated with shorter heating durations required for cytotoxicity.
  • Developed and validated a CASP8 activity indicator for monitoring system function.

Conclusions:

  • The developed thermogenetic system enables heat-inducible programmed cell death via ELP-mediated CASP8 activation.
  • This system offers precise spatiotemporal control over cellular processes, with potential applications in research and therapy.
  • Optical heating combined with thermometry provides a method for targeted single-cell manipulation.