Source 1primary paper2020Angewandte Chemie International Edition
Toolkit/caging strategy for crRNA
caging strategy for crRNA
Also known as: vitamin E-caged crRNA strategy, vitamin E modification
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The caging strategy for crRNA is a photoregulated CRISPR/Cas9 control method in which vitamin E is coupled to the 5' terminus of crRNA through a photolabile linker. In the caged state, the modified crRNA is reported to suppress target DNA association while preserving Cas9/crRNA/tracrRNA ribonucleoprotein assembly, and light irradiation restores genome-editing activity.
Usefulness & Problems
Why this is useful
This method provides optical control over CRISPR/Cas9-mediated editing, enabling light-dependent activation of genome editing in human cells. The source literature states that it could support spatiotemporal photoregulation of CRISPR/Cas9 activity.
Source:
as well as gene knockdown of EGFP expression in EGFP stably expressing cells
Source:
This new caging strategy for crRNA could provide new methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
Problem solved
It addresses the problem of controlling when and where CRISPR/Cas9 editing occurs by keeping crRNA function inactive until light exposure. The reported application is photoregulated editing of VEGFA and light-activated knockdown of EGFP expression in cells.
Source:
as well as gene knockdown of EGFP expression in EGFP stably expressing cells
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete method used to build, optimize, or evolve an engineered system.
Techniques
No technique tags yet.
Target processes
editingInput: Light
Implementation Constraints
The method uses a crRNA chemically modified at the 5' terminus with vitamin E via a photolabile linker. It is implemented within the CRISPR/Cas9 system using crRNA and tracrRNA, and activation requires light irradiation; the provided evidence does not further detail construct preparation or delivery conditions.
The supplied evidence comes from a single 2020 study and provides limited quantitative performance detail. The available text does not specify irradiation wavelength, uncaging efficiency, editing efficiency, off-target effects, or validation beyond the reported cellular examples.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells.
Upon light irradiation, vitamin E-caged crRNA was successfully activated to achieve light-induced genome editing of vascular endothelial cell-growth factor A (VEGFA) in human cells through a T7E1 assay and Sanger sequencing
Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells.
Upon light irradiation, vitamin E-caged crRNA was successfully activated to achieve light-induced genome editing of vascular endothelial cell-growth factor A (VEGFA) in human cells through a T7E1 assay and Sanger sequencing
Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells.
Upon light irradiation, vitamin E-caged crRNA was successfully activated to achieve light-induced genome editing of vascular endothelial cell-growth factor A (VEGFA) in human cells through a T7E1 assay and Sanger sequencing
Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells.
Upon light irradiation, vitamin E-caged crRNA was successfully activated to achieve light-induced genome editing of vascular endothelial cell-growth factor A (VEGFA) in human cells through a T7E1 assay and Sanger sequencing
Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells.
Upon light irradiation, vitamin E-caged crRNA was successfully activated to achieve light-induced genome editing of vascular endothelial cell-growth factor A (VEGFA) in human cells through a T7E1 assay and Sanger sequencing
Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells.
Upon light irradiation, vitamin E-caged crRNA was successfully activated to achieve light-induced genome editing of vascular endothelial cell-growth factor A (VEGFA) in human cells through a T7E1 assay and Sanger sequencing
Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells.
Upon light irradiation, vitamin E-caged crRNA was successfully activated to achieve light-induced genome editing of vascular endothelial cell-growth factor A (VEGFA) in human cells through a T7E1 assay and Sanger sequencing
Light-activated vitamin E-caged crRNA enables knockdown of EGFP expression in EGFP stably expressing cells.
as well as gene knockdown of EGFP expression in EGFP stably expressing cells
Light-activated vitamin E-caged crRNA enables knockdown of EGFP expression in EGFP stably expressing cells.
as well as gene knockdown of EGFP expression in EGFP stably expressing cells
Light-activated vitamin E-caged crRNA enables knockdown of EGFP expression in EGFP stably expressing cells.
as well as gene knockdown of EGFP expression in EGFP stably expressing cells
Light-activated vitamin E-caged crRNA enables knockdown of EGFP expression in EGFP stably expressing cells.
as well as gene knockdown of EGFP expression in EGFP stably expressing cells
Light-activated vitamin E-caged crRNA enables knockdown of EGFP expression in EGFP stably expressing cells.
as well as gene knockdown of EGFP expression in EGFP stably expressing cells
Light-activated vitamin E-caged crRNA enables knockdown of EGFP expression in EGFP stably expressing cells.
as well as gene knockdown of EGFP expression in EGFP stably expressing cells
Light-activated vitamin E-caged crRNA enables knockdown of EGFP expression in EGFP stably expressing cells.
as well as gene knockdown of EGFP expression in EGFP stably expressing cells
This crRNA caging strategy could provide methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This new caging strategy for crRNA could provide new methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This crRNA caging strategy could provide methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This new caging strategy for crRNA could provide new methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This crRNA caging strategy could provide methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This new caging strategy for crRNA could provide new methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This crRNA caging strategy could provide methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This new caging strategy for crRNA could provide new methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This crRNA caging strategy could provide methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This new caging strategy for crRNA could provide new methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This crRNA caging strategy could provide methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This new caging strategy for crRNA could provide new methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This crRNA caging strategy could provide methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This new caging strategy for crRNA could provide new methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
Coupling vitamin E and a photolabile linker at the 5' terminus of crRNA inactivates the CRISPR/Cas9 system.
Here, we synthesized a novel light-controlled crRNA by coupling vitamin E and a photolabile linker at the 5' terminus to inactivate the CRISPR/Cas9 system.
Coupling vitamin E and a photolabile linker at the 5' terminus of crRNA inactivates the CRISPR/Cas9 system.
Here, we synthesized a novel light-controlled crRNA by coupling vitamin E and a photolabile linker at the 5' terminus to inactivate the CRISPR/Cas9 system.
Coupling vitamin E and a photolabile linker at the 5' terminus of crRNA inactivates the CRISPR/Cas9 system.
Here, we synthesized a novel light-controlled crRNA by coupling vitamin E and a photolabile linker at the 5' terminus to inactivate the CRISPR/Cas9 system.
Coupling vitamin E and a photolabile linker at the 5' terminus of crRNA inactivates the CRISPR/Cas9 system.
Here, we synthesized a novel light-controlled crRNA by coupling vitamin E and a photolabile linker at the 5' terminus to inactivate the CRISPR/Cas9 system.
Coupling vitamin E and a photolabile linker at the 5' terminus of crRNA inactivates the CRISPR/Cas9 system.
Here, we synthesized a novel light-controlled crRNA by coupling vitamin E and a photolabile linker at the 5' terminus to inactivate the CRISPR/Cas9 system.
Coupling vitamin E and a photolabile linker at the 5' terminus of crRNA inactivates the CRISPR/Cas9 system.
Here, we synthesized a novel light-controlled crRNA by coupling vitamin E and a photolabile linker at the 5' terminus to inactivate the CRISPR/Cas9 system.
Coupling vitamin E and a photolabile linker at the 5' terminus of crRNA inactivates the CRISPR/Cas9 system.
Here, we synthesized a novel light-controlled crRNA by coupling vitamin E and a photolabile linker at the 5' terminus to inactivate the CRISPR/Cas9 system.
The vitamin E modification does not affect formation of Cas9/crRNA/tracrRNA ribonucleoprotein complexes.
The vitamin E modification did not affect ribonucleoprotein (RNP) formation of Cas9/crRNA/tracrRNA complexes
The vitamin E modification does not affect formation of Cas9/crRNA/tracrRNA ribonucleoprotein complexes.
The vitamin E modification did not affect ribonucleoprotein (RNP) formation of Cas9/crRNA/tracrRNA complexes
The vitamin E modification does not affect formation of Cas9/crRNA/tracrRNA ribonucleoprotein complexes.
The vitamin E modification did not affect ribonucleoprotein (RNP) formation of Cas9/crRNA/tracrRNA complexes
The vitamin E modification does not affect formation of Cas9/crRNA/tracrRNA ribonucleoprotein complexes.
The vitamin E modification did not affect ribonucleoprotein (RNP) formation of Cas9/crRNA/tracrRNA complexes
The vitamin E modification does not affect formation of Cas9/crRNA/tracrRNA ribonucleoprotein complexes.
The vitamin E modification did not affect ribonucleoprotein (RNP) formation of Cas9/crRNA/tracrRNA complexes
The vitamin E modification does not affect formation of Cas9/crRNA/tracrRNA ribonucleoprotein complexes.
The vitamin E modification did not affect ribonucleoprotein (RNP) formation of Cas9/crRNA/tracrRNA complexes
The vitamin E modification does not affect formation of Cas9/crRNA/tracrRNA ribonucleoprotein complexes.
The vitamin E modification did not affect ribonucleoprotein (RNP) formation of Cas9/crRNA/tracrRNA complexes
The vitamin E modification inhibits association of the Cas9/crRNA/tracrRNA ribonucleoprotein complex with target DNA.
but did inhibit the association of RNP with the target DNA
The vitamin E modification inhibits association of the Cas9/crRNA/tracrRNA ribonucleoprotein complex with target DNA.
but did inhibit the association of RNP with the target DNA
The vitamin E modification inhibits association of the Cas9/crRNA/tracrRNA ribonucleoprotein complex with target DNA.
but did inhibit the association of RNP with the target DNA
The vitamin E modification inhibits association of the Cas9/crRNA/tracrRNA ribonucleoprotein complex with target DNA.
but did inhibit the association of RNP with the target DNA
The vitamin E modification inhibits association of the Cas9/crRNA/tracrRNA ribonucleoprotein complex with target DNA.
but did inhibit the association of RNP with the target DNA
The vitamin E modification inhibits association of the Cas9/crRNA/tracrRNA ribonucleoprotein complex with target DNA.
but did inhibit the association of RNP with the target DNA
The vitamin E modification inhibits association of the Cas9/crRNA/tracrRNA ribonucleoprotein complex with target DNA.
but did inhibit the association of RNP with the target DNA
Approval Evidence
1 source1 linked approval claimfirst-pass slug caging-strategy-for-crrna
This new caging strategy for crRNA could provide new methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
Source:
capabilitysupports
This crRNA caging strategy could provide methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
This new caging strategy for crRNA could provide new methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
Source:
Comparisons
Source-backed strengths
The strategy was reported to enable light-activated genome editing of VEGFA in human cells and knockdown of EGFP in EGFP-stably expressing cells. Its design is described as maintaining Cas9/crRNA/tracrRNA complex assembly while suppressing target DNA association before irradiation.
Ranked Citations
- 1.