Source 1primary paper2020Nature Communications
Toolkit/CRISPRoff
CRISPRoff
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
CRISPRoff is an engineering method for light-induced degradation of single-guide RNA (sgRNA) that inactivates the CRISPR ribonucleoprotein. It enables spatiotemporal attenuation of CRISPR-Cas9 genome editing in cells through selective illumination.
Usefulness & Problems
Why this is useful
CRISPRoff is useful for imposing optical control over CRISPR editing activity in cells. The reported system allows titratable editing efficiency and spatial patterning via selective illumination, supporting experiments that require temporal and positional restriction of genome editing.
Source:
Here we show that light-induced inactivation of ribonucleoprotein attenuates genome editing within cells
Problem solved
This method addresses the problem of how to turn down or locally restrict CRISPR-Cas9 activity after delivery in cells. According to the cited study, it does so by using light-induced sgRNA degradation to attenuate genome editing.
Source:
This uncontrolled reaction can lead to unintended consequences including off-target editing and chromosomal translocations.
Published Workflows
Objective: Combine CRISPR-based endogenous gene regulation with synthetic Notch signaling to record state-specific cell-cell interactions and intentionally influence the outcomes of those interactions in engineered human cells.
Why it works: The workflow is presented as effective because artificial receptors provide defined input-to-output linkages for cell decision-making, while CRISPR-based strategies allow endogenous gene programs to be genetically or epigenetically manipulated to control which interactions are recorded and how cells respond.
epigenetic manipulation of endogenous gene expressionartificial receptor-mediated cell-cell signalingconditional relay of cell-state-dependent interaction signalsCRISPR-based genetic manipulationCRISPR-based epigenetic manipulationsynthetic receptor engineering
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete method used to build, optimize, or evolve an engineered system.
Mechanisms
attenuation of genome editingcrispr ribonucleoprotein inactivationDegradationlight-induced inactivation of the crispr ribonucleoproteinlight-induced sgrna degradationTechniques
No technique tags yet.
Target processes
degradationeditinglocalizationInput: Light
Implementation Constraints
The available evidence indicates that the method relies on light input and is built around sgRNA degradation to inactivate the CRISPR ribonucleoprotein in cells. The supplied material does not provide construct architecture, required cofactors, expression strategy, or illumination parameters.
The provided evidence is limited to a single source and does not specify the photochemical design, illumination wavelength, degradation machinery, or quantitative performance metrics. Independent replication, organismal breadth, and compatibility across Cas systems or delivery formats are not established by the supplied evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Source 2primary paper2024MED
Ranked Claims
Signal-competent interactions mediated through the platform can direct differentiation of human embryonic stem cells toward pre-selected fates based on assigned synNotch outputs.
Further, such signal-competent interactions can be used to direct differentiation of human embryonic stem cells toward pre-selected fates based on assigned synNotch outputs.
The combined CRISPR-based and synthetic Notch approach enables conditional recording of interactions between human cells based on expression of a state-specific marker in a subset of cells.
Our approach gives rise to the ability to conditionally record interactions between human cells, where the record of engagement depends on expression of a state-specific marker of a subset of cells in a population.
CRISPR-based manipulation of native gene expression profiles can bias outcomes of cell engagement histories in a targeted manner.
We also implemented CRISPR-based manipulation of native gene expression profiles to bias outcomes of cell engagement histories in a targeted manner.
CRISPRoff allows titratable levels of editing efficiency and spatial patterning via selective illumination.
allows for titratable levels of editing efficiency and spatial patterning via selective illumination
CRISPRoff allows titratable levels of editing efficiency and spatial patterning via selective illumination.
allows for titratable levels of editing efficiency and spatial patterning via selective illumination
CRISPRoff allows titratable levels of editing efficiency and spatial patterning via selective illumination.
allows for titratable levels of editing efficiency and spatial patterning via selective illumination
CRISPRoff allows titratable levels of editing efficiency and spatial patterning via selective illumination.
allows for titratable levels of editing efficiency and spatial patterning via selective illumination
CRISPRoff allows titratable levels of editing efficiency and spatial patterning via selective illumination.
allows for titratable levels of editing efficiency and spatial patterning via selective illumination
CRISPRoff allows titratable levels of editing efficiency and spatial patterning via selective illumination.
allows for titratable levels of editing efficiency and spatial patterning via selective illumination
CRISPRoff allows titratable levels of editing efficiency and spatial patterning via selective illumination.
allows for titratable levels of editing efficiency and spatial patterning via selective illumination
Light-induced inactivation of the CRISPR ribonucleoprotein attenuates genome editing within cells.
Here we show that light-induced inactivation of ribonucleoprotein attenuates genome editing within cells
Light-induced inactivation of the CRISPR ribonucleoprotein attenuates genome editing within cells.
Here we show that light-induced inactivation of ribonucleoprotein attenuates genome editing within cells
Light-induced inactivation of the CRISPR ribonucleoprotein attenuates genome editing within cells.
Here we show that light-induced inactivation of ribonucleoprotein attenuates genome editing within cells
Light-induced inactivation of the CRISPR ribonucleoprotein attenuates genome editing within cells.
Here we show that light-induced inactivation of ribonucleoprotein attenuates genome editing within cells
Light-induced inactivation of the CRISPR ribonucleoprotein attenuates genome editing within cells.
Here we show that light-induced inactivation of ribonucleoprotein attenuates genome editing within cells
Light-induced inactivation of the CRISPR ribonucleoprotein attenuates genome editing within cells.
Here we show that light-induced inactivation of ribonucleoprotein attenuates genome editing within cells
Light-induced inactivation of the CRISPR ribonucleoprotein attenuates genome editing within cells.
Here we show that light-induced inactivation of ribonucleoprotein attenuates genome editing within cells
CRISPRoff is a method for light-induced degradation of sgRNA.
To address this, we develop a method for light-induced degradation of sgRNA termed CRISPRoff.
CRISPRoff is a method for light-induced degradation of sgRNA.
To address this, we develop a method for light-induced degradation of sgRNA termed CRISPRoff.
CRISPRoff is a method for light-induced degradation of sgRNA.
To address this, we develop a method for light-induced degradation of sgRNA termed CRISPRoff.
CRISPRoff is a method for light-induced degradation of sgRNA.
To address this, we develop a method for light-induced degradation of sgRNA termed CRISPRoff.
CRISPRoff is a method for light-induced degradation of sgRNA.
To address this, we develop a method for light-induced degradation of sgRNA termed CRISPRoff.
CRISPRoff is a method for light-induced degradation of sgRNA.
To address this, we develop a method for light-induced degradation of sgRNA termed CRISPRoff.
CRISPRoff is a method for light-induced degradation of sgRNA.
To address this, we develop a method for light-induced degradation of sgRNA termed CRISPRoff.
Uncontrolled CRISPR-Cas9 activity after cellular introduction can lead to off-target editing and chromosomal translocations.
This uncontrolled reaction can lead to unintended consequences including off-target editing and chromosomal translocations.
Uncontrolled CRISPR-Cas9 activity after cellular introduction can lead to off-target editing and chromosomal translocations.
This uncontrolled reaction can lead to unintended consequences including off-target editing and chromosomal translocations.
Uncontrolled CRISPR-Cas9 activity after cellular introduction can lead to off-target editing and chromosomal translocations.
This uncontrolled reaction can lead to unintended consequences including off-target editing and chromosomal translocations.
Uncontrolled CRISPR-Cas9 activity after cellular introduction can lead to off-target editing and chromosomal translocations.
This uncontrolled reaction can lead to unintended consequences including off-target editing and chromosomal translocations.
Uncontrolled CRISPR-Cas9 activity after cellular introduction can lead to off-target editing and chromosomal translocations.
This uncontrolled reaction can lead to unintended consequences including off-target editing and chromosomal translocations.
Uncontrolled CRISPR-Cas9 activity after cellular introduction can lead to off-target editing and chromosomal translocations.
This uncontrolled reaction can lead to unintended consequences including off-target editing and chromosomal translocations.
Uncontrolled CRISPR-Cas9 activity after cellular introduction can lead to off-target editing and chromosomal translocations.
This uncontrolled reaction can lead to unintended consequences including off-target editing and chromosomal translocations.
Approval Evidence
2 sources6 linked approval claimsfirst-pass slug crisproff
Use of CRISPRoff and synthetic Notch to modulate and relay endogenous gene expression programs in engineered cells.
Source:
To address this, we develop a method for light-induced degradation of sgRNA termed CRISPRoff.
Source:
capabilitysupports
The combined CRISPR-based and synthetic Notch approach enables conditional recording of interactions between human cells based on expression of a state-specific marker in a subset of cells.
Our approach gives rise to the ability to conditionally record interactions between human cells, where the record of engagement depends on expression of a state-specific marker of a subset of cells in a population.
Source:
modulationsupports
CRISPR-based manipulation of native gene expression profiles can bias outcomes of cell engagement histories in a targeted manner.
We also implemented CRISPR-based manipulation of native gene expression profiles to bias outcomes of cell engagement histories in a targeted manner.
Source:
control capabilitysupports
CRISPRoff allows titratable levels of editing efficiency and spatial patterning via selective illumination.
allows for titratable levels of editing efficiency and spatial patterning via selective illumination
Source:
functional effectsupports
Light-induced inactivation of the CRISPR ribonucleoprotein attenuates genome editing within cells.
Here we show that light-induced inactivation of ribonucleoprotein attenuates genome editing within cells
Source:
method developmentsupports
CRISPRoff is a method for light-induced degradation of sgRNA.
To address this, we develop a method for light-induced degradation of sgRNA termed CRISPRoff.
Source:
problem statementsupports
Uncontrolled CRISPR-Cas9 activity after cellular introduction can lead to off-target editing and chromosomal translocations.
This uncontrolled reaction can lead to unintended consequences including off-target editing and chromosomal translocations.
Source:
Comparisons
Source-backed strengths
The source reports that CRISPRoff provides spatiotemporal control of CRISPR editing in cells. It also supports titratable levels of editing efficiency and spatial patterning under selective illumination, indicating controllable optical modulation of CRISPR activity.
Ranked Citations
- 1.
- 2.