Source 1primary paper2019Nature Communications
Toolkit/ligand-activated and ligand-deactivated sgRNAs
ligand-activated and ligand-deactivated sgRNAs
Also known as: ligand-activated sgRNAs, ligand-deactivated sgRNAs, sgRNA, sgRNAs, single guide RNA, single guide RNA (sgRNA)
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Ligand-activated and ligand-deactivated sgRNAs are engineered single-guide RNAs that place CRISPR-Cas9-based gene repression under small-molecule control. In the reported 2019 Nature Communications study, sgRNA function was activated or deactivated in a dose-dependent manner by two different ligands, with regulation acting directly at the level of each target-specific sgRNA.
Usefulness & Problems
Why this is useful
These sgRNAs are useful for chemically controlling CRISPR-Cas9 repression without changing the target-specific Cas9 protein component. Because control is encoded in individual sgRNAs, the system supports differential and opposing temporal regulation of multiple genes within the same framework.
Source:
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
Source:
We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands.
Source:
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
Source:
We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands.
Problem solved
This tool addresses the problem of achieving target-specific, small-molecule-responsive control over CRISPR-Cas9-based gene repression. It specifically helps solve the need for independently tunable activation or deactivation of different sgRNAs, enabling temporal control across multiple gene targets.
Source:
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
Source:
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
Problem links
Need controllable genome or transcript editing
DerivedLigand-activated and ligand-deactivated sgRNAs are engineered single-guide RNAs that place CRISPR-Cas9-based gene repression under small-molecule control. In the reported 2019 Nature Communications study, sgRNA function was activated or deactivated in a dose-dependent manner by two different ligands, with regulation acting directly at the level of each target-specific sgRNA.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level RNA part used inside a larger architecture that realizes a mechanism.
Mechanisms
chemical activation of crispr-cas9-based gene repressionchemical deactivation of crispr-cas9-based gene repressiondose-dependent ligand responsesmall-molecule-dependent allosteric switching of sgrna activityTechniques
No technique tags yet.
Target processes
editingInput: Chemical
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: regulatorswitch architecture: uncaging
Implementation requires engineered sgRNAs designed to be ligand responsive and used in a CRISPR-Cas9 gene repression context. The available evidence indicates that regulation is encoded directly in each target-specific sgRNA, but it does not provide construct architecture, sequence design rules, or delivery details.
The supplied evidence supports chemical control of CRISPR-Cas9-based gene repression, but it does not establish performance for genome editing or other Cas9 outputs. The evidence set is limited to a single cited study and does not provide detailed quantitative performance metrics, ligand identities, or broad cross-system validation.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Source 2primary paper2018
Ranked Claims
Because the system acts directly on each target-specific sgRNA, it enables applications requiring differential and opposing temporal control of multiple genes.
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
Because the system acts directly on each target-specific sgRNA, it enables applications requiring differential and opposing temporal control of multiple genes.
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
Because the system acts directly on each target-specific sgRNA, it enables applications requiring differential and opposing temporal control of multiple genes.
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
Because the system acts directly on each target-specific sgRNA, it enables applications requiring differential and opposing temporal control of multiple genes.
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
Because the system acts directly on each target-specific sgRNA, it enables applications requiring differential and opposing temporal control of multiple genes.
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
Because the system acts directly on each target-specific sgRNA, it enables applications requiring differential and opposing temporal control of multiple genes.
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
Because the system acts directly on each target-specific sgRNA, it enables applications requiring differential and opposing temporal control of multiple genes.
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
CRISPR-Cas9-based gene repression can be activated or deactivated in a dose-dependent manner in response to two different small-molecule ligands.
We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands.
dynamic range 10 fold
CRISPR-Cas9-based gene repression can be activated or deactivated in a dose-dependent manner in response to two different small-molecule ligands.
We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands.
dynamic range 10 fold
CRISPR-Cas9-based gene repression can be activated or deactivated in a dose-dependent manner in response to two different small-molecule ligands.
We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands.
dynamic range 10 fold
CRISPR-Cas9-based gene repression can be activated or deactivated in a dose-dependent manner in response to two different small-molecule ligands.
We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands.
dynamic range 10 fold
CRISPR-Cas9-based gene repression can be activated or deactivated in a dose-dependent manner in response to two different small-molecule ligands.
We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands.
dynamic range 10 fold
CRISPR-Cas9-based gene repression can be activated or deactivated in a dose-dependent manner in response to two different small-molecule ligands.
We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands.
dynamic range 10 fold
CRISPR-Cas9-based gene repression can be activated or deactivated in a dose-dependent manner in response to two different small-molecule ligands.
We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands.
dynamic range 10 fold
Because the system acts directly on each target-specific sgRNA, it enables differential and opposing temporal control of multiple genes.
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
CRISPR-Cas9-based gene repression can be activated or deactivated in a dose-dependent manner in response to two different small-molecule ligands using the reported sgRNA system.
We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands.
dynamic range 10 fold
Approval Evidence
2 sources4 linked approval claimsfirst-pass slug ligand-activated-and-ligand-deactivated-sgrnas
Controlling CRISPR-Cas9 with ligand-activated and ligand-deactivated sgRNAs
Source:
Controlling CRISPR-Cas9 with ligand-activated and ligand-deactivated sgRNAs
Source:
application scopesupports
Because the system acts directly on each target-specific sgRNA, it enables applications requiring differential and opposing temporal control of multiple genes.
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
Source:
functional effectsupports
CRISPR-Cas9-based gene repression can be activated or deactivated in a dose-dependent manner in response to two different small-molecule ligands.
We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands.
Source:
application scopesupports
Because the system acts directly on each target-specific sgRNA, it enables differential and opposing temporal control of multiple genes.
Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
Source:
functional effectsupports
CRISPR-Cas9-based gene repression can be activated or deactivated in a dose-dependent manner in response to two different small-molecule ligands using the reported sgRNA system.
We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands.
Source:
Comparisons
Source-backed strengths
The reported system provides both ligand-activated and ligand-deactivated modes of sgRNA control, expanding the range of possible regulatory logic. It was shown to respond in a dose-dependent manner to two different small-molecule ligands, and its action at individual target-specific sgRNAs enables differential and opposing control of multiple genes.
Compared with aptazyme-embedded guide RNAs
ligand-activated and ligand-deactivated sgRNAs and aptazyme-embedded guide RNAs address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing; same primary input modality: chemical
Strengths here: appears more independently replicated; looks easier to implement in practice.
Compared with photo-sensitive circular gRNAs
ligand-activated and ligand-deactivated sgRNAs 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
Strengths here: appears more independently replicated; looks easier to implement in practice.
Compared with small molecule regulated sgRNAs
ligand-activated and ligand-deactivated sgRNAs and small molecule regulated sgRNAs address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing
Strengths here: appears more independently replicated; looks easier to implement in practice.
Ranked Citations
- 1.