Source 1primary paper2024UDSpace Institutional Repository (University of Delaware)
Toolkit/toehold-gated guide RNA
toehold-gated guide RNA
Also known as: thgRNA
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Toehold-gated guide RNA (thgRNA) is a synthetic riboregulatory guide RNA class that controls CRISPR/Cas9 activity in response to RNA inputs. Available evidence indicates that endogenous RNA transcripts can trigger thgRNA to activate Cas9 functions, supporting autonomous RNA-responsive control of genome-targeting activity.
Usefulness & Problems
Why this is useful
thgRNA is useful as an RNA-responsive control element for coupling CRISPR/Cas9 function to cellular transcript states. The supplied evidence specifically supports its use for autonomous control schemes in which endogenous RNA transcripts gate Cas9 activity.
Problem solved
thgRNA addresses the problem of making Cas9 activity conditional on the presence of specific RNA signals rather than constitutively active. This enables dynamic control architectures in which endogenous transcripts serve as internal triggers for CRISPR function.
Problem links
Need controllable genome or transcript editing
DerivedToehold-gated guide RNA (thgRNA) is a synthetic riboregulatory guide RNA class that controls CRISPR/Cas9 activity in response to RNA inputs. Available evidence indicates that endogenous RNA transcripts can trigger thgRNA to activate Cas9 functions, supporting autonomous RNA-responsive control of genome-targeting activity.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level RNA part used inside a larger architecture that realizes a mechanism.
Mechanisms
conditional activation of cas9 by endogenous or programmed rna inputsconditional activation of cas9 by endogenous rna inputsrna-triggered conformational gating of guide rna activitytoehold-mediated rna sensingTechniques
No technique tags yet.
Target processes
editingImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: regulator
Implementation is supported at the conceptual level as a guide RNA engineered with a toehold-gated regulatory architecture that responds to endogenous RNA transcripts. The supplied evidence does not specify Cas9 variant, scaffold design, expression system, delivery method, or construct constraints.
The provided evidence is sparse and does not include sequence design rules, activation kinetics, dynamic range, off-target effects, or organism-specific validation details. Claims about sensing full-length mRNA, multiplexing, and minimal cross-talk are not directly supported by the supplied excerpts.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Endogenous RNA transcripts can trigger toehold-gated guide RNA to activate Cas9 functions, enabling autonomous control elements in dynamic control schemes.
The versatility of this platform was demonstrated by the use of endogenous RNA transcripts as triggers to activate Cas9 functions, allowing thgRNA to be used as an autonomous control elements in dynamic control schemes.
Endogenous RNA transcripts can trigger toehold-gated guide RNA to activate Cas9 functions, enabling autonomous control elements in dynamic control schemes.
The versatility of this platform was demonstrated by the use of endogenous RNA transcripts as triggers to activate Cas9 functions, allowing thgRNA to be used as an autonomous control elements in dynamic control schemes.
Endogenous RNA transcripts can trigger toehold-gated guide RNA to activate Cas9 functions, enabling autonomous control elements in dynamic control schemes.
The versatility of this platform was demonstrated by the use of endogenous RNA transcripts as triggers to activate Cas9 functions, allowing thgRNA to be used as an autonomous control elements in dynamic control schemes.
Endogenous RNA transcripts can trigger toehold-gated guide RNA to activate Cas9 functions, enabling autonomous control elements in dynamic control schemes.
The versatility of this platform was demonstrated by the use of endogenous RNA transcripts as triggers to activate Cas9 functions, allowing thgRNA to be used as an autonomous control elements in dynamic control schemes.
Endogenous RNA transcripts can trigger toehold-gated guide RNA to activate Cas9 functions, enabling autonomous control elements in dynamic control schemes.
The versatility of this platform was demonstrated by the use of endogenous RNA transcripts as triggers to activate Cas9 functions, allowing thgRNA to be used as an autonomous control elements in dynamic control schemes.
Endogenous RNA transcripts can trigger toehold-gated guide RNA to activate Cas9 functions, enabling autonomous control elements in dynamic control schemes.
The versatility of this platform was demonstrated by the use of endogenous RNA transcripts as triggers to activate Cas9 functions, allowing thgRNA to be used as an autonomous control elements in dynamic control schemes.
Endogenous RNA transcripts can trigger toehold-gated guide RNA to activate Cas9 functions, enabling autonomous control elements in dynamic control schemes.
The versatility of this platform was demonstrated by the use of endogenous RNA transcripts as triggers to activate Cas9 functions, allowing thgRNA to be used as an autonomous control elements in dynamic control schemes.
A synthetic extracellular sensing circuit can exploit pre-existing membrane receptors through input-induced reconstitution of a native signaling peptide.
we envisioned a synthetic extracellular sensing circuit that can exploit pre-existing membrane receptors by input-induced reconstitution of native signaling peptide
A synthetic extracellular sensing circuit can exploit pre-existing membrane receptors through input-induced reconstitution of a native signaling peptide.
we envisioned a synthetic extracellular sensing circuit that can exploit pre-existing membrane receptors by input-induced reconstitution of native signaling peptide
A synthetic extracellular sensing circuit can exploit pre-existing membrane receptors through input-induced reconstitution of a native signaling peptide.
we envisioned a synthetic extracellular sensing circuit that can exploit pre-existing membrane receptors by input-induced reconstitution of native signaling peptide
A synthetic extracellular sensing circuit can exploit pre-existing membrane receptors through input-induced reconstitution of a native signaling peptide.
we envisioned a synthetic extracellular sensing circuit that can exploit pre-existing membrane receptors by input-induced reconstitution of native signaling peptide
A synthetic extracellular sensing circuit can exploit pre-existing membrane receptors through input-induced reconstitution of a native signaling peptide.
we envisioned a synthetic extracellular sensing circuit that can exploit pre-existing membrane receptors by input-induced reconstitution of native signaling peptide
A synthetic extracellular sensing circuit can exploit pre-existing membrane receptors through input-induced reconstitution of a native signaling peptide.
we envisioned a synthetic extracellular sensing circuit that can exploit pre-existing membrane receptors by input-induced reconstitution of native signaling peptide
A synthetic extracellular sensing circuit can exploit pre-existing membrane receptors through input-induced reconstitution of a native signaling peptide.
we envisioned a synthetic extracellular sensing circuit that can exploit pre-existing membrane receptors by input-induced reconstitution of native signaling peptide
Intein-mediated reactions were adapted to reconstitute the yeast mating pheromone alpha-factor and use the associated yeast mating pathway to direct cellular responses.
The biochemical basis of such a sensing circuit was established by adapting intein-mediated reactions to reconstitute the well-known yeast mating pheromone peptide, α-factor, and exploiting the associated yeast mating pathway to direct cellular responses.
Intein-mediated reactions were adapted to reconstitute the yeast mating pheromone alpha-factor and use the associated yeast mating pathway to direct cellular responses.
The biochemical basis of such a sensing circuit was established by adapting intein-mediated reactions to reconstitute the well-known yeast mating pheromone peptide, α-factor, and exploiting the associated yeast mating pathway to direct cellular responses.
Intein-mediated reactions were adapted to reconstitute the yeast mating pheromone alpha-factor and use the associated yeast mating pathway to direct cellular responses.
The biochemical basis of such a sensing circuit was established by adapting intein-mediated reactions to reconstitute the well-known yeast mating pheromone peptide, α-factor, and exploiting the associated yeast mating pathway to direct cellular responses.
Intein-mediated reactions were adapted to reconstitute the yeast mating pheromone alpha-factor and use the associated yeast mating pathway to direct cellular responses.
The biochemical basis of such a sensing circuit was established by adapting intein-mediated reactions to reconstitute the well-known yeast mating pheromone peptide, α-factor, and exploiting the associated yeast mating pathway to direct cellular responses.
Intein-mediated reactions were adapted to reconstitute the yeast mating pheromone alpha-factor and use the associated yeast mating pathway to direct cellular responses.
The biochemical basis of such a sensing circuit was established by adapting intein-mediated reactions to reconstitute the well-known yeast mating pheromone peptide, α-factor, and exploiting the associated yeast mating pathway to direct cellular responses.
Intein-mediated reactions were adapted to reconstitute the yeast mating pheromone alpha-factor and use the associated yeast mating pathway to direct cellular responses.
The biochemical basis of such a sensing circuit was established by adapting intein-mediated reactions to reconstitute the well-known yeast mating pheromone peptide, α-factor, and exploiting the associated yeast mating pathway to direct cellular responses.
Intein-mediated reactions were adapted to reconstitute the yeast mating pheromone alpha-factor and use the associated yeast mating pathway to direct cellular responses.
The biochemical basis of such a sensing circuit was established by adapting intein-mediated reactions to reconstitute the well-known yeast mating pheromone peptide, α-factor, and exploiting the associated yeast mating pathway to direct cellular responses.
Toehold-gated guide RNA can be programmed to respond to a wide variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
The synthetic riboregulators can be programmed to respond to a very large variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
Toehold-gated guide RNA can be programmed to respond to a wide variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
The synthetic riboregulators can be programmed to respond to a very large variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
Toehold-gated guide RNA can be programmed to respond to a wide variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
The synthetic riboregulators can be programmed to respond to a very large variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
Toehold-gated guide RNA can be programmed to respond to a wide variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
The synthetic riboregulators can be programmed to respond to a very large variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
Toehold-gated guide RNA can be programmed to respond to a wide variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
The synthetic riboregulators can be programmed to respond to a very large variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
Toehold-gated guide RNA can be programmed to respond to a wide variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
The synthetic riboregulators can be programmed to respond to a very large variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
Toehold-gated guide RNA can be programmed to respond to a wide variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
The synthetic riboregulators can be programmed to respond to a very large variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
Approval Evidence
1 source2 linked approval claimsfirst-pass slug toehold-gated-guide-rna
a new class of riboregulators, termed toehold-gated guide RNA
Source:
autonomous controlsupports
Endogenous RNA transcripts can trigger toehold-gated guide RNA to activate Cas9 functions, enabling autonomous control elements in dynamic control schemes.
The versatility of this platform was demonstrated by the use of endogenous RNA transcripts as triggers to activate Cas9 functions, allowing thgRNA to be used as an autonomous control elements in dynamic control schemes.
Source:
programmabilitysupports
Toehold-gated guide RNA can be programmed to respond to a wide variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
The synthetic riboregulators can be programmed to respond to a very large variety of RNA sequences, including full-length mRNA, and control CRISPR/Cas9 activities for multiplexed gene regulation in E. coli with minimal cross-talk.
Source:
Comparisons
Source-backed strengths
The evidence supports that thgRNA constitutes a distinct class of riboregulators and that endogenous RNA can activate its associated Cas9 function. This indicates programmability at the RNA-input level, but the supplied source does not provide quantitative performance metrics or comparative benchmarks.
Compared with antisense oligonucleotides
toehold-gated guide RNA and antisense oligonucleotides address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing
Strengths here: looks easier to implement in practice.
Compared with photo-sensitive circular gRNAs
toehold-gated guide RNA 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: looks easier to implement in practice.
Compared with synthetic riboswitches
toehold-gated guide RNA and synthetic riboswitches address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing
Relative tradeoffs: appears more independently replicated; looks easier to implement in practice.
Ranked Citations
- 1.