Source 1primary paper2019
Toolkit/PIF6-CreN
PIF6-CreN
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
PIF6-CreN is the PIF6-fused N-terminal Cre fragment used in the CreLite optogenetic recombinase system. In the presence of phycocyanobilin (PCB), 660 nm red light induces association with PhyB-CreC, reconstituting Cre recombinase activity for light-controlled loxP recombination in developing zebrafish embryos.
Usefulness & Problems
Why this is useful
This component enables red-light control of Cre/loxP recombination when paired with PhyB-CreC and PCB. In zebrafish embryos, this system supported Cre-dependent multi-spectral cell labeling in heart, skeletal muscle, and epithelium after red-light exposure.
Source:
Red-light exposure of transgenic zebrafish embryos harboring a Cre-dependent multi-color fluorescent protein reporter ( ubi:zebrabow ) injected with CreLite mRNAs and PCB, resulted in Cre activity as measured by the generation of multi-spectral cell labeling in various tissues, including heart, skeletal muscle and epithelium.
Source:
Here we present CreLite, an optogenetically-controlled Cre system using red light in developing zebrafish embryos.
Source:
We show that CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
Problem solved
PIF6-CreN helps solve the problem of making Cre recombinase activity conditional on an external optical input rather than constitutively active. The reported design disables Cre by splitting it and restores activity only when red light and PCB drive interaction with the complementary PhyB-CreC fusion.
Source:
Red-light exposure of transgenic zebrafish embryos harboring a Cre-dependent multi-color fluorescent protein reporter ( ubi:zebrabow ) injected with CreLite mRNAs and PCB, resulted in Cre activity as measured by the generation of multi-spectral cell labeling in various tissues, including heart, skeletal muscle and epithelium.
Source:
We show that CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Techniques
No technique tags yet.
Target processes
recombinationInput: Light
Implementation Constraints
The reported implementation involved injection of CreLite mRNAs and PCB into transgenic zebrafish embryos carrying a Cre-dependent multi-color fluorescent reporter. Activity depended on 660 nm red-light exposure and on co-presence of the PhyB-CreC fusion partner, consistent with a multi-component construct design based on split Cre fragments fused to light-inducible binding partners.
PIF6-CreN is not functional as a standalone reagent and requires the complementary PhyB-CreC component plus PCB to restore Cre activity. The supplied evidence is limited to a single reported system in developing zebrafish embryos, with no independent replication or broader organismal validation provided here.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
In transgenic zebrafish embryos carrying a Cre-dependent multi-color fluorescent reporter, red-light exposure after injection with CreLite mRNAs and PCB resulted in Cre activity measured by multi-spectral cell labeling in heart, skeletal muscle, and epithelium.
Red-light exposure of transgenic zebrafish embryos harboring a Cre-dependent multi-color fluorescent protein reporter ( ubi:zebrabow ) injected with CreLite mRNAs and PCB, resulted in Cre activity as measured by the generation of multi-spectral cell labeling in various tissues, including heart, skeletal muscle and epithelium.
In transgenic zebrafish embryos carrying a Cre-dependent multi-color fluorescent reporter, red-light exposure after injection with CreLite mRNAs and PCB resulted in Cre activity measured by multi-spectral cell labeling in heart, skeletal muscle, and epithelium.
Red-light exposure of transgenic zebrafish embryos harboring a Cre-dependent multi-color fluorescent protein reporter ( ubi:zebrabow ) injected with CreLite mRNAs and PCB, resulted in Cre activity as measured by the generation of multi-spectral cell labeling in various tissues, including heart, skeletal muscle and epithelium.
In transgenic zebrafish embryos carrying a Cre-dependent multi-color fluorescent reporter, red-light exposure after injection with CreLite mRNAs and PCB resulted in Cre activity measured by multi-spectral cell labeling in heart, skeletal muscle, and epithelium.
Red-light exposure of transgenic zebrafish embryos harboring a Cre-dependent multi-color fluorescent protein reporter ( ubi:zebrabow ) injected with CreLite mRNAs and PCB, resulted in Cre activity as measured by the generation of multi-spectral cell labeling in various tissues, including heart, skeletal muscle and epithelium.
In transgenic zebrafish embryos carrying a Cre-dependent multi-color fluorescent reporter, red-light exposure after injection with CreLite mRNAs and PCB resulted in Cre activity measured by multi-spectral cell labeling in heart, skeletal muscle, and epithelium.
Red-light exposure of transgenic zebrafish embryos harboring a Cre-dependent multi-color fluorescent protein reporter ( ubi:zebrabow ) injected with CreLite mRNAs and PCB, resulted in Cre activity as measured by the generation of multi-spectral cell labeling in various tissues, including heart, skeletal muscle and epithelium.
In transgenic zebrafish embryos carrying a Cre-dependent multi-color fluorescent reporter, red-light exposure after injection with CreLite mRNAs and PCB resulted in Cre activity measured by multi-spectral cell labeling in heart, skeletal muscle, and epithelium.
Red-light exposure of transgenic zebrafish embryos harboring a Cre-dependent multi-color fluorescent protein reporter ( ubi:zebrabow ) injected with CreLite mRNAs and PCB, resulted in Cre activity as measured by the generation of multi-spectral cell labeling in various tissues, including heart, skeletal muscle and epithelium.
In transgenic zebrafish embryos carrying a Cre-dependent multi-color fluorescent reporter, red-light exposure after injection with CreLite mRNAs and PCB resulted in Cre activity measured by multi-spectral cell labeling in heart, skeletal muscle, and epithelium.
Red-light exposure of transgenic zebrafish embryos harboring a Cre-dependent multi-color fluorescent protein reporter ( ubi:zebrabow ) injected with CreLite mRNAs and PCB, resulted in Cre activity as measured by the generation of multi-spectral cell labeling in various tissues, including heart, skeletal muscle and epithelium.
In transgenic zebrafish embryos carrying a Cre-dependent multi-color fluorescent reporter, red-light exposure after injection with CreLite mRNAs and PCB resulted in Cre activity measured by multi-spectral cell labeling in heart, skeletal muscle, and epithelium.
Red-light exposure of transgenic zebrafish embryos harboring a Cre-dependent multi-color fluorescent protein reporter ( ubi:zebrabow ) injected with CreLite mRNAs and PCB, resulted in Cre activity as measured by the generation of multi-spectral cell labeling in various tissues, including heart, skeletal muscle and epithelium.
CreLite disables Cre by splitting Cre and fusing the inactive halves to the red light-inducible binding partners PhyB and PIF6.
Cre activity is disabled by splitting Cre and fusing the inactive halves with the Arabidopsis thaliana red light-inducible binding partners, PhyB and PIF6.
CreLite disables Cre by splitting Cre and fusing the inactive halves to the red light-inducible binding partners PhyB and PIF6.
Cre activity is disabled by splitting Cre and fusing the inactive halves with the Arabidopsis thaliana red light-inducible binding partners, PhyB and PIF6.
CreLite disables Cre by splitting Cre and fusing the inactive halves to the red light-inducible binding partners PhyB and PIF6.
Cre activity is disabled by splitting Cre and fusing the inactive halves with the Arabidopsis thaliana red light-inducible binding partners, PhyB and PIF6.
CreLite disables Cre by splitting Cre and fusing the inactive halves to the red light-inducible binding partners PhyB and PIF6.
Cre activity is disabled by splitting Cre and fusing the inactive halves with the Arabidopsis thaliana red light-inducible binding partners, PhyB and PIF6.
CreLite disables Cre by splitting Cre and fusing the inactive halves to the red light-inducible binding partners PhyB and PIF6.
Cre activity is disabled by splitting Cre and fusing the inactive halves with the Arabidopsis thaliana red light-inducible binding partners, PhyB and PIF6.
CreLite disables Cre by splitting Cre and fusing the inactive halves to the red light-inducible binding partners PhyB and PIF6.
Cre activity is disabled by splitting Cre and fusing the inactive halves with the Arabidopsis thaliana red light-inducible binding partners, PhyB and PIF6.
CreLite disables Cre by splitting Cre and fusing the inactive halves to the red light-inducible binding partners PhyB and PIF6.
Cre activity is disabled by splitting Cre and fusing the inactive halves with the Arabidopsis thaliana red light-inducible binding partners, PhyB and PIF6.
Red light at 660 nm in the presence of PCB brings PhyB-CreC and PIF6-CreN together to restore Cre activity.
Upon exposure to red light (660 nm) illumination, the PhyB-CreC and PIF6-CreN fusion proteins come together in the presence of PCB to restore Cre activity.
activation wavelength 660 nm
Red light at 660 nm in the presence of PCB brings PhyB-CreC and PIF6-CreN together to restore Cre activity.
Upon exposure to red light (660 nm) illumination, the PhyB-CreC and PIF6-CreN fusion proteins come together in the presence of PCB to restore Cre activity.
activation wavelength 660 nm
Red light at 660 nm in the presence of PCB brings PhyB-CreC and PIF6-CreN together to restore Cre activity.
Upon exposure to red light (660 nm) illumination, the PhyB-CreC and PIF6-CreN fusion proteins come together in the presence of PCB to restore Cre activity.
activation wavelength 660 nm
Red light at 660 nm in the presence of PCB brings PhyB-CreC and PIF6-CreN together to restore Cre activity.
Upon exposure to red light (660 nm) illumination, the PhyB-CreC and PIF6-CreN fusion proteins come together in the presence of PCB to restore Cre activity.
activation wavelength 660 nm
Red light at 660 nm in the presence of PCB brings PhyB-CreC and PIF6-CreN together to restore Cre activity.
Upon exposure to red light (660 nm) illumination, the PhyB-CreC and PIF6-CreN fusion proteins come together in the presence of PCB to restore Cre activity.
activation wavelength 660 nm
Red light at 660 nm in the presence of PCB brings PhyB-CreC and PIF6-CreN together to restore Cre activity.
Upon exposure to red light (660 nm) illumination, the PhyB-CreC and PIF6-CreN fusion proteins come together in the presence of PCB to restore Cre activity.
activation wavelength 660 nm
Red light at 660 nm in the presence of PCB brings PhyB-CreC and PIF6-CreN together to restore Cre activity.
Upon exposure to red light (660 nm) illumination, the PhyB-CreC and PIF6-CreN fusion proteins come together in the presence of PCB to restore Cre activity.
activation wavelength 660 nm
CreLite is an optogenetically controlled Cre system that uses red light in developing zebrafish embryos.
Here we present CreLite, an optogenetically-controlled Cre system using red light in developing zebrafish embryos.
CreLite is an optogenetically controlled Cre system that uses red light in developing zebrafish embryos.
Here we present CreLite, an optogenetically-controlled Cre system using red light in developing zebrafish embryos.
CreLite is an optogenetically controlled Cre system that uses red light in developing zebrafish embryos.
Here we present CreLite, an optogenetically-controlled Cre system using red light in developing zebrafish embryos.
CreLite is an optogenetically controlled Cre system that uses red light in developing zebrafish embryos.
Here we present CreLite, an optogenetically-controlled Cre system using red light in developing zebrafish embryos.
CreLite is an optogenetically controlled Cre system that uses red light in developing zebrafish embryos.
Here we present CreLite, an optogenetically-controlled Cre system using red light in developing zebrafish embryos.
CreLite is an optogenetically controlled Cre system that uses red light in developing zebrafish embryos.
Here we present CreLite, an optogenetically-controlled Cre system using red light in developing zebrafish embryos.
CreLite is an optogenetically controlled Cre system that uses red light in developing zebrafish embryos.
Here we present CreLite, an optogenetically-controlled Cre system using red light in developing zebrafish embryos.
CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
We show that CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
We show that CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
We show that CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
We show that CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
We show that CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
We show that CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
We show that CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development.
Approval Evidence
1 source2 linked approval claimsfirst-pass slug pif6-cren
Upon exposure to red light (660 nm) illumination, the PhyB-CreC and PIF6-CreN fusion proteins come together in the presence of PCB to restore Cre activity.
Source:
mechanismsupports
CreLite disables Cre by splitting Cre and fusing the inactive halves to the red light-inducible binding partners PhyB and PIF6.
Cre activity is disabled by splitting Cre and fusing the inactive halves with the Arabidopsis thaliana red light-inducible binding partners, PhyB and PIF6.
Source:
mechanismsupports
Red light at 660 nm in the presence of PCB brings PhyB-CreC and PIF6-CreN together to restore Cre activity.
Upon exposure to red light (660 nm) illumination, the PhyB-CreC and PIF6-CreN fusion proteins come together in the presence of PCB to restore Cre activity.
Source:
Comparisons
Source-backed strengths
The system uses 660 nm red light to trigger Cre reconstitution, providing an optically gated recombination input with defined wavelength dependence in the reported study. Functional activity was demonstrated in vivo in developing zebrafish embryos using a Cre-dependent fluorescent reporter, with labeling observed in multiple tissues.
Ranked Citations
- 1.