Scientific Reports (Dec 2023)

Optogenetic induction of caspase-8 mediated apoptosis by employing Arabidopsis cryptochrome 2

  • Weiliang Mo,
  • Shengzhong Su,
  • Ruige Shang,
  • Liang Yang,
  • Xuelai Zhao,
  • Chengfeng Wu,
  • Zhenming Yang,
  • He Zhang,
  • Liuming Wu,
  • Yibo Liu,
  • Yun He,
  • Ruipeng Zhang,
  • Zecheng Zuo

DOI
https://doi.org/10.1038/s41598-023-50561-y
Journal volume & issue
Vol. 13, no. 1
pp. 1 – 8

Abstract

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Abstract Apoptosis, a programmed cell death mechanism, is a regulatory process controlling cell proliferation as cells undergo demise. Caspase-8 serves as a pivotal apoptosis-inducing factor that initiates the death receptor-mediated apoptosis pathway. In this investigation, we have devised an optogenetic method to swiftly modulate caspase-8 activation in response to blue light. The cornerstone of our optogenetic tool relies on the PHR domain of Arabidopsis thaliana cryptochrome 2, which self-oligomerizes upon exposure to blue light. In this study, we have developed two optogenetic approaches for rapidly controlling caspase-8 activation in response to blue light in cellular systems. The first strategy, denoted as Opto-Casp8-V1, entails the fusion expression of the Arabidopsis blue light receptor CRY2 N-terminal PHR domain with caspase-8. The second strategy, referred to as Opto-Casp8-V2, involves the independent fusion expression of caspase-8 with the PHR domain and the CRY2 blue light-interacting protein CIB1 N-terminal CIB1N. Upon induction with blue light, PHR undergoes aggregation, leading to caspase-8 aggregation. Additionally, the blue light-dependent interaction between PHR and CIB1N also results in caspase-8 aggregation. We have validated these strategies in both HEK293T and HeLa cells. The findings reveal that both strategies are capable of inducing apoptosis, with Opto-Casp8-V2 demonstrating significantly superior efficiency compared to Opto-Casp8-V1.