Toolkit Items

FUN-LOV is a fungal light-oxygen-voltage optogenetic switch for yeast built from Neurospora crassa photoreceptors WC-1 and VVD. It uses the photon-regulated interaction of these components to drive light-dependent target gene activation, including GPD1 and ADH1 expression in a wine yeast strain.

Cry2 is a blue-light photoreceptor cryptochrome from Arabidopsis used as a light-responsive multi-component optogenetic switch. The supplied evidence supports blue-light-dependent photoactivation linked to regulation of transcription factor control and to CRY2 degradation.

iLID/SspB is a blue-light-inducible heterodimerization system built from an engineered iLID module and the SspB binding partner. It is used to reversibly recruit proteins in cells for control of localization and signaling, including membrane recruitment, neurotrophin receptor construction, microtubule plus-end targeting, and perturbation of small GTPase pathways.

BcLOV4 is a blue-light-responsive photoreceptor from Botrytis cinerea that functions as a single-component optogenetic module for rapid plasma membrane recruitment. Illumination drives cytosol-to-membrane translocation through direct protein-lipid electrostatic interaction, and fused cargo such as RhoA GTPase or RhoA-activating GEFs can be relocalized to the membrane with light.

OptoRAF1 is a blue light-responsive optogenetic RAF1 system built on the CRY2/CIB1 dimerizer pair. It reversibly activates the RAF/MEK/ERK pathway by recruiting RAF1 to the plasma membrane.

Opto-Rho1DN is a multi-component optogenetic switch that inhibits Rho1 by light-dependent recruitment of a dominant-negative Rho1 construct to the plasma membrane. The listed components are CIBN-pmGFP and CRY2-Rho1DN.

Applications of microscopy and plant tissue culture have included elucidation of growth and development processes, detection of in vitro-induced physiological disorders as well as subcellular localization using fluorescent protein probes.

The light-switchable condensate system is a genetically encoded, multi-component platform for blue light-controlled organization of functional cargoes in Escherichia coli. It couples a condensation-enabling scaffold to cargo proteins through the iLID/SspB light-responsive interaction pair to dynamically regulate cargo localization.

BcLOV4-ARHGEF11 is a single-transgene optogenetic fusion in which the upstream RhoA activator ARHGEF11 is fused to BcLOV4. It enables light-inducible, spatiotemporally precise control of RhoA signaling through dynamic membrane localization without requiring a separate protein binding partner.

The BcLOV4-RhoA optogenetic fusion is a single-transgene light-responsive construct in which RhoA GTPase, or its upstream activator ARHGEF11, is fused to BcLOV4. It enables spatiotemporally precise optical control of RhoA signaling and associated cytoskeletal and mechanotransductive responses without requiring a separate protein binding partner for dynamic membrane localization.

The CRY2-talin/CIBN-CAAX optogenetic plasma membrane recruitment system is a blue-light-responsive two-component switch that fuses Arabidopsis cryptochrome 2 to the N terminus of full-length talin and anchors the N-terminal cryptochrome-interacting basic helix-loop-helix domain to the plasma membrane with a CAAX motif. In Chinese hamster ovary cells and endothelial cells, 450 nm illumination recruits talin to the plasma membrane and promotes activation of β3 integrins, including αIIbβ3- and αVβ3-associated complexes.

Light-inducible TrkA activation strategies comprise four engineered optical designs for activating TrkA signaling without nerve growth factor. The reported approaches use light to drive plasma membrane recruitment and homo-interaction of the intracellular domain of TrkA, recapitulating native NGF/TrkA-associated functions.

optoTGFBRs is an optogenetic multi-component switch developed to control TGF-β signaling with light. It enables precise spatiotemporal regulation of the pathway and has been used to drive selective and sequential activation in single cells.

p21-CRY2/CIB1 is a blue-light-responsive optogenetic p21 system built by fusing p21 to the cryptochrome 2/CIBN switch components. It was used to control p21 subcellular localization and nuclear function and increased the fraction of cells arrested in G1 phase.

PiGM-Iq is a light-controlled multi-component switch for Photo-induced Modulation of Gα protein signaling that inhibits Gαq. The reported system produced potent and selective inhibition of Gαq signaling and was applied to perturb signaling-dependent neural development and behavior.

The re-engineered MLKL membrane recruitment tool is a single-component optogenetic system derived from MLKL that preserves light-mediated plasma membrane recruitment while blocking MLKL-associated cell-killing activity. It is intended to modulate protein function through light-controlled localization at the plasma membrane.

Single-component optogenetic tools were created to control RhoA GTPase signaling with light. The reported system does not require protein binding partners and enables inducible RhoA-mediated cytoskeletal activation with downstream YAP nuclear localization and YAP-TEAD mechanotranscription.

p21-LINuS is a light-controllable p21 construct created by fusing p21 to the AsLOV-based light-inducible nuclear localization signal (LINuS). It was used to optogenetically control p21 subcellular localization and nuclear function, increasing the fraction of cells arrested in G1 phase under blue LED illumination.

Chromophore-assisted laser or light inactivation (CALI) has been employed as a promising technique to achieve spatiotemporal knockdown or loss-of-function of target molecules in situ.