Source 1primary paper2020Angewandte Chemie International Edition
Toolkit/light-controlled crRNA
light-controlled crRNA
Also known as: photolabile crRNA, vitamin E-caged crRNA
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Light-controlled crRNA is a chemically modified CRISPR guide RNA in which vitamin E is attached to the 5' terminus through a photolabile linker, creating a caged crRNA that inactivates CRISPR/Cas9 until light exposure. Upon irradiation, this design restores CRISPR/Cas9 activity and supports genome editing of VEGFA and knockdown of EGFP expression in human cells.
Usefulness & Problems
Why this is useful
This tool provides optical control over CRISPR/Cas9 activity, enabling light-dependent activation of genome editing rather than constitutive guide function. The reported strategy is useful for pursuing spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.
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 how to keep CRISPR/Cas9 inactive before a chosen time and then activate editing with light. The reported implementation specifically uses a 5' vitamin E photolabile cage on crRNA to suppress activity until irradiation.
Source:
as well as gene knockdown of EGFP expression in EGFP stably expressing cells
Problem links
Need controllable genome or transcript editing
DerivedLight-controlled crRNA is a chemically modified CRISPR guide RNA in which vitamin E is attached to the 5' terminus through a photolabile linker, creating a caged crRNA that inactivates CRISPR/Cas9 until light exposure. Upon irradiation, this design restores CRISPR/Cas9 activity and supports genome editing of VEGFA and knockdown of EGFP expression in human cells.
Need precise spatiotemporal control with light input
DerivedLight-controlled crRNA is a chemically modified CRISPR guide RNA in which vitamin E is attached to the 5' terminus through a photolabile linker, creating a caged crRNA that inactivates CRISPR/Cas9 until light exposure. Upon irradiation, this design restores CRISPR/Cas9 activity and supports genome editing of VEGFA and knockdown of EGFP expression in human cells.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level RNA part used inside a larger architecture that realizes a mechanism.
Target processes
editingInput: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: spectral hardware requirementoperating role: regulatorswitch architecture: cleavage
The reported construct is a crRNA chemically modified at the 5' terminus with vitamin E via a photolabile linker. The available evidence supports use with CRISPR/Cas9 in human cells, but it does not provide practical details on delivery format, Cas9 expression system, or illumination parameters.
The supplied evidence comes from a single 2020 study and supports only a limited set of demonstrated targets and applications. The evidence provided does not specify irradiation wavelength, uncaging kinetics, editing efficiency, off-target effects, or performance across additional cell types or organisms.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Observations
successMammalian Cell Lineapplication demohuman
T7E1 assay and Sanger sequencing
Inferred from claim c4 during normalization. Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells. Derived from claim c4. Quoted text: 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
Source:
successMammalian Cell Lineapplication demohuman
T7E1 assay and Sanger sequencing
Inferred from claim c4 during normalization. Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells. Derived from claim c4. Quoted text: 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
Source:
successMammalian Cell Lineapplication demohuman
T7E1 assay and Sanger sequencing
Inferred from claim c4 during normalization. Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells. Derived from claim c4. Quoted text: 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
Source:
successMammalian Cell Lineapplication demohuman
T7E1 assay and Sanger sequencing
Inferred from claim c4 during normalization. Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells. Derived from claim c4. Quoted text: 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
Source:
successMammalian Cell Lineapplication demohuman
T7E1 assay and Sanger sequencing
Inferred from claim c4 during normalization. Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells. Derived from claim c4. Quoted text: 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
Source:
successMammalian Cell Lineapplication demohuman
T7E1 assay and Sanger sequencing
Inferred from claim c4 during normalization. Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells. Derived from claim c4. Quoted text: 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
Source:
successMammalian Cell Lineapplication demohuman
T7E1 assay and Sanger sequencing
Inferred from claim c4 during normalization. Light irradiation activates vitamin E-caged crRNA to enable genome editing of VEGFA in human cells. Derived from claim c4. Quoted text: 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
Source:
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 source5 linked approval claimsfirst-pass slug light-controlled-crrna
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.
Source:
activation effectsupports
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
Source:
applicationsupports
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
Source:
mechanismsupports
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.
Source:
mechanismsupports
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
Source:
mechanismsupports
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
Source:
Comparisons
Source-backed strengths
The tool was reported to enable light-activated genome editing of VEGFA in human cells and light-activated knockdown of EGFP expression in EGFP-stably expressing cells. Its design is chemically defined, based on 5' terminal coupling of vitamin E through a photolabile linker, and directly links activation to light exposure.
light-controlled crRNA and auxiliary photocleavable oligodeoxyribonucleotides complementary to crRNA address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing; shared mechanisms: photocleavage; same primary input modality: light
Compared with caged guide RNA
light-controlled crRNA and caged guide RNA address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing; shared mechanisms: photocleavage; same primary input modality: light
Compared with photo-sensitive circular gRNAs
light-controlled crRNA and photo-sensitive circular gRNAs address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing; shared mechanisms: photocleavage; same primary input modality: light
Ranked Citations
- 1.