Source 1primary paper2018OakTrust (Texas A&M University Libraries)
Toolkit/OptoORAI1
OptoORAI1
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
OptoORAI1 is a photoswitchable CRAC channel engineered from ORAI1 by insertion of a LOV2 photosensory domain into an ORAI1 loop region. In this design, LOV2 functions as an allosteric light-responsive switch that opens the channel, enabling optical control of calcium signaling.
Usefulness & Problems
Why this is useful
OptoORAI1 belongs to the OptoCRAC toolkit, which enables remote and precise control of calcium signaling with high spatial and temporal resolution. This makes the system useful for manipulating Ca2+-dependent cellular programs, including NFAT-linked transcriptional outputs demonstrated for OptoCRAC tools.
Source:
our single-component OptoCRAC tools provide new opportunities to remotely and precisely control the Ca^2+ signaling at high spatial and temporal resolution
Source:
We have successfully demonstrated the use of OptoCRAC to photo-tune Ca^2+/NFAT-dependent gene expression, as well as transcriptional reprogramming of endogenous genes when coupled with the CRISPR/Cas9 genome editing technique.
Problem solved
OptoORAI1 addresses the problem of controlling CRAC channel activity and downstream calcium signaling with light rather than conventional chemical or constitutive inputs. The cited work positions this as a way to achieve precise spatiotemporal regulation of Ca2+-dependent signaling and gene regulation.
Source:
our single-component OptoCRAC tools provide new opportunities to remotely and precisely control the Ca^2+ signaling at high spatial and temporal resolution
Source:
We have successfully demonstrated the use of OptoCRAC to photo-tune Ca^2+/NFAT-dependent gene expression, as well as transcriptional reprogramming of endogenous genes when coupled with the CRISPR/Cas9 genome editing technique.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Target processes
recombinationselectionInput: Light
Implementation Constraints
The construct was generated by inserting LOV2 into a loop region of ORAI1 so that the photosensory domain acts as an allosteric switch on channel opening. The same source also reports development of cpLOV2 variants by circular permutation to create new interfaces for caging protein function, but the evidence does not specify whether a cpLOV2 variant was required in the final OptoORAI1 construct.
The supplied evidence does not provide OptoORAI1-specific quantitative performance metrics such as activation wavelength, kinetics, dynamic range, leak, or calcium conductance. Independent replication and validation outside the cited source are not provided in the evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
OptoCRAC tools enable remote and precise control of calcium signaling with high spatial and temporal resolution.
our single-component OptoCRAC tools provide new opportunities to remotely and precisely control the Ca^2+ signaling at high spatial and temporal resolution
OptoCRAC tools enable remote and precise control of calcium signaling with high spatial and temporal resolution.
our single-component OptoCRAC tools provide new opportunities to remotely and precisely control the Ca^2+ signaling at high spatial and temporal resolution
OptoCRAC tools enable remote and precise control of calcium signaling with high spatial and temporal resolution.
our single-component OptoCRAC tools provide new opportunities to remotely and precisely control the Ca^2+ signaling at high spatial and temporal resolution
OptoCRAC tools enable remote and precise control of calcium signaling with high spatial and temporal resolution.
our single-component OptoCRAC tools provide new opportunities to remotely and precisely control the Ca^2+ signaling at high spatial and temporal resolution
OptoCRAC tools enable remote and precise control of calcium signaling with high spatial and temporal resolution.
our single-component OptoCRAC tools provide new opportunities to remotely and precisely control the Ca^2+ signaling at high spatial and temporal resolution
OptoCRAC tools enable remote and precise control of calcium signaling with high spatial and temporal resolution.
our single-component OptoCRAC tools provide new opportunities to remotely and precisely control the Ca^2+ signaling at high spatial and temporal resolution
OptoCRAC tools enable remote and precise control of calcium signaling with high spatial and temporal resolution.
our single-component OptoCRAC tools provide new opportunities to remotely and precisely control the Ca^2+ signaling at high spatial and temporal resolution
OptoCRAC was successfully used to photo-tune Ca2+/NFAT-dependent gene expression and to reprogram endogenous gene transcription when coupled with CRISPR/Cas9.
We have successfully demonstrated the use of OptoCRAC to photo-tune Ca^2+/NFAT-dependent gene expression, as well as transcriptional reprogramming of endogenous genes when coupled with the CRISPR/Cas9 genome editing technique.
OptoCRAC was successfully used to photo-tune Ca2+/NFAT-dependent gene expression and to reprogram endogenous gene transcription when coupled with CRISPR/Cas9.
We have successfully demonstrated the use of OptoCRAC to photo-tune Ca^2+/NFAT-dependent gene expression, as well as transcriptional reprogramming of endogenous genes when coupled with the CRISPR/Cas9 genome editing technique.
OptoCRAC was successfully used to photo-tune Ca2+/NFAT-dependent gene expression and to reprogram endogenous gene transcription when coupled with CRISPR/Cas9.
We have successfully demonstrated the use of OptoCRAC to photo-tune Ca^2+/NFAT-dependent gene expression, as well as transcriptional reprogramming of endogenous genes when coupled with the CRISPR/Cas9 genome editing technique.
OptoCRAC was successfully used to photo-tune Ca2+/NFAT-dependent gene expression and to reprogram endogenous gene transcription when coupled with CRISPR/Cas9.
We have successfully demonstrated the use of OptoCRAC to photo-tune Ca^2+/NFAT-dependent gene expression, as well as transcriptional reprogramming of endogenous genes when coupled with the CRISPR/Cas9 genome editing technique.
OptoCRAC was successfully used to photo-tune Ca2+/NFAT-dependent gene expression and to reprogram endogenous gene transcription when coupled with CRISPR/Cas9.
We have successfully demonstrated the use of OptoCRAC to photo-tune Ca^2+/NFAT-dependent gene expression, as well as transcriptional reprogramming of endogenous genes when coupled with the CRISPR/Cas9 genome editing technique.
OptoCRAC was successfully used to photo-tune Ca2+/NFAT-dependent gene expression and to reprogram endogenous gene transcription when coupled with CRISPR/Cas9.
We have successfully demonstrated the use of OptoCRAC to photo-tune Ca^2+/NFAT-dependent gene expression, as well as transcriptional reprogramming of endogenous genes when coupled with the CRISPR/Cas9 genome editing technique.
OptoCRAC was successfully used to photo-tune Ca2+/NFAT-dependent gene expression and to reprogram endogenous gene transcription when coupled with CRISPR/Cas9.
We have successfully demonstrated the use of OptoCRAC to photo-tune Ca^2+/NFAT-dependent gene expression, as well as transcriptional reprogramming of endogenous genes when coupled with the CRISPR/Cas9 genome editing technique.
cpLOV2 variants were developed through circular permutation to create new interfaces for caging protein function.
we developed a series of engineered LOV2 variants (cpLOV2) through circular permutation. cpLOV2 creates new interfaces to cage protein function
OptoORAI1 was generated by inserting LOV2 into the loop region of ORAI1 so that LOV2 acts as an allosteric switch to open the channel.
To generate OptoORAI1, LOV2 was inserted into the loop region of ORAI1 and thus acted as an allosteric switch to induce structural rearrangement within ORAI1 to open the channel.
OptoORAI1 was generated by inserting LOV2 into the loop region of ORAI1 so that LOV2 acts as an allosteric switch to open the channel.
To generate OptoORAI1, LOV2 was inserted into the loop region of ORAI1 and thus acted as an allosteric switch to induce structural rearrangement within ORAI1 to open the channel.
OptoORAI1 was generated by inserting LOV2 into the loop region of ORAI1 so that LOV2 acts as an allosteric switch to open the channel.
To generate OptoORAI1, LOV2 was inserted into the loop region of ORAI1 and thus acted as an allosteric switch to induce structural rearrangement within ORAI1 to open the channel.
OptoORAI1 was generated by inserting LOV2 into the loop region of ORAI1 so that LOV2 acts as an allosteric switch to open the channel.
To generate OptoORAI1, LOV2 was inserted into the loop region of ORAI1 and thus acted as an allosteric switch to induce structural rearrangement within ORAI1 to open the channel.
OptoORAI1 was generated by inserting LOV2 into the loop region of ORAI1 so that LOV2 acts as an allosteric switch to open the channel.
To generate OptoORAI1, LOV2 was inserted into the loop region of ORAI1 and thus acted as an allosteric switch to induce structural rearrangement within ORAI1 to open the channel.
OptoORAI1 was generated by inserting LOV2 into the loop region of ORAI1 so that LOV2 acts as an allosteric switch to open the channel.
To generate OptoORAI1, LOV2 was inserted into the loop region of ORAI1 and thus acted as an allosteric switch to induce structural rearrangement within ORAI1 to open the channel.
OptoORAI1 was generated by inserting LOV2 into the loop region of ORAI1 so that LOV2 acts as an allosteric switch to open the channel.
To generate OptoORAI1, LOV2 was inserted into the loop region of ORAI1 and thus acted as an allosteric switch to induce structural rearrangement within ORAI1 to open the channel.
OptoSTIM1 was engineered by combining the STIM1 SOAR region with the LOV2 domain.
OptoSTIM1 was engineered by combining STIM1-ORAI1 activation region (SOAR) of STIM1 with the light-reactive light-oxygen-voltage (LOV2) domain.
OptoSTIM1 was engineered by combining the STIM1 SOAR region with the LOV2 domain.
OptoSTIM1 was engineered by combining STIM1-ORAI1 activation region (SOAR) of STIM1 with the light-reactive light-oxygen-voltage (LOV2) domain.
OptoSTIM1 was engineered by combining the STIM1 SOAR region with the LOV2 domain.
OptoSTIM1 was engineered by combining STIM1-ORAI1 activation region (SOAR) of STIM1 with the light-reactive light-oxygen-voltage (LOV2) domain.
OptoSTIM1 was engineered by combining the STIM1 SOAR region with the LOV2 domain.
OptoSTIM1 was engineered by combining STIM1-ORAI1 activation region (SOAR) of STIM1 with the light-reactive light-oxygen-voltage (LOV2) domain.
OptoSTIM1 was engineered by combining the STIM1 SOAR region with the LOV2 domain.
OptoSTIM1 was engineered by combining STIM1-ORAI1 activation region (SOAR) of STIM1 with the light-reactive light-oxygen-voltage (LOV2) domain.
OptoSTIM1 was engineered by combining the STIM1 SOAR region with the LOV2 domain.
OptoSTIM1 was engineered by combining STIM1-ORAI1 activation region (SOAR) of STIM1 with the light-reactive light-oxygen-voltage (LOV2) domain.
OptoSTIM1 was engineered by combining the STIM1 SOAR region with the LOV2 domain.
OptoSTIM1 was engineered by combining STIM1-ORAI1 activation region (SOAR) of STIM1 with the light-reactive light-oxygen-voltage (LOV2) domain.
Randomized screening and optimization identified an OptoORAI1 variant with high light-induced calcium response change and no noticeable dark activity.
Through several rounds of randomized screening and optimization, we identified one OptoORAI1 variant exhibiting a high dynamic change in the light-induced Ca^2+ response without noticeable dark activity.
Randomized screening and optimization identified an OptoORAI1 variant with high light-induced calcium response change and no noticeable dark activity.
Through several rounds of randomized screening and optimization, we identified one OptoORAI1 variant exhibiting a high dynamic change in the light-induced Ca^2+ response without noticeable dark activity.
Randomized screening and optimization identified an OptoORAI1 variant with high light-induced calcium response change and no noticeable dark activity.
Through several rounds of randomized screening and optimization, we identified one OptoORAI1 variant exhibiting a high dynamic change in the light-induced Ca^2+ response without noticeable dark activity.
Randomized screening and optimization identified an OptoORAI1 variant with high light-induced calcium response change and no noticeable dark activity.
Through several rounds of randomized screening and optimization, we identified one OptoORAI1 variant exhibiting a high dynamic change in the light-induced Ca^2+ response without noticeable dark activity.
Randomized screening and optimization identified an OptoORAI1 variant with high light-induced calcium response change and no noticeable dark activity.
Through several rounds of randomized screening and optimization, we identified one OptoORAI1 variant exhibiting a high dynamic change in the light-induced Ca^2+ response without noticeable dark activity.
Randomized screening and optimization identified an OptoORAI1 variant with high light-induced calcium response change and no noticeable dark activity.
Through several rounds of randomized screening and optimization, we identified one OptoORAI1 variant exhibiting a high dynamic change in the light-induced Ca^2+ response without noticeable dark activity.
Randomized screening and optimization identified an OptoORAI1 variant with high light-induced calcium response change and no noticeable dark activity.
Through several rounds of randomized screening and optimization, we identified one OptoORAI1 variant exhibiting a high dynamic change in the light-induced Ca^2+ response without noticeable dark activity.
Approval Evidence
1 source2 linked approval claimsfirst-pass slug optoorai1
or ORAI1 (OptoORAI1) to generate photoswitchable CRAC channels
Source:
engineering strategysupports
OptoORAI1 was generated by inserting LOV2 into the loop region of ORAI1 so that LOV2 acts as an allosteric switch to open the channel.
To generate OptoORAI1, LOV2 was inserted into the loop region of ORAI1 and thus acted as an allosteric switch to induce structural rearrangement within ORAI1 to open the channel.
Source:
screening outcomesupports
Randomized screening and optimization identified an OptoORAI1 variant with high light-induced calcium response change and no noticeable dark activity.
Through several rounds of randomized screening and optimization, we identified one OptoORAI1 variant exhibiting a high dynamic change in the light-induced Ca^2+ response without noticeable dark activity.
Source:
Comparisons
Source-backed strengths
The reported design directly converts ORAI1 into a photoswitchable CRAC channel through LOV2 insertion, providing an allosteric route to light-gated channel opening. The broader OptoCRAC platform was shown to support photo-tuning of Ca2+/NFAT-dependent gene expression and reprogramming of endogenous gene transcription when coupled with CRISPR/Cas9.
Source:
we developed a series of engineered LOV2 variants (cpLOV2) through circular permutation. cpLOV2 creates new interfaces to cage protein function
Source:
To generate OptoORAI1, LOV2 was inserted into the loop region of ORAI1 and thus acted as an allosteric switch to induce structural rearrangement within ORAI1 to open the channel.
Source:
OptoSTIM1 was engineered by combining STIM1-ORAI1 activation region (SOAR) of STIM1 with the light-reactive light-oxygen-voltage (LOV2) domain.
Ranked Citations
- 1.