Optogenetic activators of apoptosis, necroptosis and pyroptosis for probing cell death dynamics and bystander cell responses

Abstract

Targeted and specific induction of cell death in individual or groups of cells holds the potential for new insights into the response of tissues or organisms to different forms of death. Here we report the development of optogenetically-controlled cell death effectors (optoCDEs), a novel class of optogenetic tools that enables light-mediated induction of three types of programmed cell death (PCD), apoptosis, pyroptosis and necroptosis, using Arabidopsis thaliana photosensitive protein Cryptochrome2. OptoCDEs enable rapid and highly specific induction of PCD in human, mouse and zebrafish cells and are suitable for a wide range of applications, such as sub-lethal cell death induction or precise elimination of single cells or cell populations in vitro and in vivo. As the proof-of-concept, we utilize optoCDEs to assess the differences in the neighboring cell response to apoptotic or necrotic PCD, revealing a new role for shingosine-1-phosphate signaling in regulating the efferocytosis of apoptotic cell by epithelia.

A role for the centrosome in regulating the rate of neuronal efferocytosis by microglia in vivo

“…Having established a leading role for BM engulfments in neuronal removal we used live reporters for microtubules (Tg ( UAS:EMTB-3xGFP );) and F-actin (Tg( UAS:mNG-UtrCH ); Shkarina et al, 2021), to determine the nature and dynamics of these extensions. We found that microglia have different types of extensions (Figure 2A).…”

“…Transgenic marker 6xUAS:lyn-tagRFPt p5E_6xUAS pME_lyn p3E_tagRFPt cry:ECFP 6xUAS:mNeonGreen-Rab5a p5E_6xUAS pME_mNeonGreen (Shkarina et al, 2021) p3E_Rab5 (Hartmann et al, 2020) cmlc2:tagRFP 6xUAS:Lamp1-mGFP p5E_6xUAS pME_Lamp1-mGFP p3E_polyA cmlc2:tagRFP 6xUAS:miRFP670-cetn4 p5E_6xUAS pME_miRFP670 p3E_cetn4 (Revenu et al, 2014) cry:mKate2…”

Revisiting the origin of interleukin 1 in anamniotes and sub-functionalization of interleukin 1 in amniotes

“…In mammals, the latter protein is cleaved by Caspase-1 downstream of the classical inflammasome cascade and IL-1 secretion [ 46 ]. Nevertheless, zebrafish Caspa and Caspb proteins are able to in vitro cleave the human GSDMD [ 47 ] and zebrafish Gasdermin E [ 22 , 48 ].…”

A fresh look at mycobacterial pathogenicity with the zebrafish host model

“…Additionally, there is a need for vesicle markers to trace the intracellular trafficking of mycobacteria and for fluorescently tagged proteins to study the activation of innate immunity signaling mechanisms in real time. Novel optogenetic tools are now being applied in zebrafish disease models and this technology opens new possibilities also for infection research, enabling local and timed activation of factors at the host‐pathogen interface (Asakawa et al., 2021 ; Formella et al., 2018 ; Shkarina et al., 2021 ). These and other technological advances will maximize the use of zebrafish as a model host and will drive research into the mechanisms of innate host defense and pathogen evasion strategies, the necessary basis for therapeutic innovations.…”