Source 1primary paper2022
Toolkit/Smart RNA guides
Smart RNA guides
Also known as: SmartGuides
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Smart RNA guides (SmartGuides) are engineered CRISPR-Cas9 guide RNAs that become active only in the presence of a specific RNA opener. They provide conditional RNA-responsive control of guide function and were reported in miRNA-responsive formats and Boolean logic circuit compositions.
Usefulness & Problems
Why this is useful
SmartGuides are useful for coupling CRISPR-Cas9 activity to endogenous or supplied RNA signals, enabling conditional genome-targeting behavior. The reported ability to operate with a panel of health-relevant miRNAs and to compose Boolean logic circuits indicates utility for programmable cellular decision-making.
Source:
We demonstrate this flexibility by operating SmartGuides activated by a panel of miRNA relevant for human health, and by composing SmartGuides in Boolean logic circuits.
Problem solved
This tool addresses the problem of making Cas9 guide activity contingent on a defined RNA input rather than constitutively active. It also addresses the design challenge of building RNA-responsive CRISPR control while keeping the spacer and opener independent for greater design flexibility.
Source:
We demonstrate this flexibility by operating SmartGuides activated by a panel of miRNA relevant for human health, and by composing SmartGuides in Boolean logic circuits.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level RNA part used inside a larger architecture that realizes a mechanism.
Techniques
Computational DesignTarget processes
No target processes tagged yet.
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: actuator
Implementation involves an engineered guide RNA used with CRISPR-Cas9 and an RNA opener that activates the guide. The available evidence indicates that spacer and opener can be designed independently, but it does not specify construct architecture, delivery method, expression system, or sequence design rules.
The supplied evidence establishes conditional RNA-triggered activation and application scope, but it does not provide quantitative performance metrics such as dynamic range, leakiness, kinetics, or editing efficiency. Independent replication and validation across multiple biological contexts are not documented in the provided evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Observations
mixedMammalian Cell Lineapplication demo
Inferred from claim c4 during normalization. In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation. Derived from claim c4. Quoted text: Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
Source:
mixedMammalian Cell Lineapplication demo
Inferred from claim c4 during normalization. In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation. Derived from claim c4. Quoted text: Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
Source:
mixedMammalian Cell Lineapplication demo
Inferred from claim c4 during normalization. In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation. Derived from claim c4. Quoted text: Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
Source:
mixedMammalian Cell Lineapplication demo
Inferred from claim c4 during normalization. In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation. Derived from claim c4. Quoted text: Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
Source:
mixedMammalian Cell Lineapplication demo
Inferred from claim c4 during normalization. In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation. Derived from claim c4. Quoted text: Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
Source:
mixedMammalian Cell Lineapplication demo
Inferred from claim c4 during normalization. In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation. Derived from claim c4. Quoted text: Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
Source:
mixedMammalian Cell Lineapplication demo
Inferred from claim c4 during normalization. In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation. Derived from claim c4. Quoted text: Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
Source:
Supporting Sources
Ranked Claims
SmartGuides were operated with a panel of miRNA relevant for human health and composed into Boolean logic circuits.
We demonstrate this flexibility by operating SmartGuides activated by a panel of miRNA relevant for human health, and by composing SmartGuides in Boolean logic circuits.
SmartGuides were operated with a panel of miRNA relevant for human health and composed into Boolean logic circuits.
We demonstrate this flexibility by operating SmartGuides activated by a panel of miRNA relevant for human health, and by composing SmartGuides in Boolean logic circuits.
SmartGuides were operated with a panel of miRNA relevant for human health and composed into Boolean logic circuits.
We demonstrate this flexibility by operating SmartGuides activated by a panel of miRNA relevant for human health, and by composing SmartGuides in Boolean logic circuits.
SmartGuides were operated with a panel of miRNA relevant for human health and composed into Boolean logic circuits.
We demonstrate this flexibility by operating SmartGuides activated by a panel of miRNA relevant for human health, and by composing SmartGuides in Boolean logic circuits.
SmartGuides were operated with a panel of miRNA relevant for human health and composed into Boolean logic circuits.
We demonstrate this flexibility by operating SmartGuides activated by a panel of miRNA relevant for human health, and by composing SmartGuides in Boolean logic circuits.
SmartGuides were operated with a panel of miRNA relevant for human health and composed into Boolean logic circuits.
We demonstrate this flexibility by operating SmartGuides activated by a panel of miRNA relevant for human health, and by composing SmartGuides in Boolean logic circuits.
SmartGuides were operated with a panel of miRNA relevant for human health and composed into Boolean logic circuits.
We demonstrate this flexibility by operating SmartGuides activated by a panel of miRNA relevant for human health, and by composing SmartGuides in Boolean logic circuits.
Smart RNA guides are conditionally activated by the presence of a specific RNA opener.
Here we report smart RNA guides that are conditionally activated by the presence of a specific RNA opener.
Smart RNA guides are conditionally activated by the presence of a specific RNA opener.
Here we report smart RNA guides that are conditionally activated by the presence of a specific RNA opener.
Smart RNA guides are conditionally activated by the presence of a specific RNA opener.
Here we report smart RNA guides that are conditionally activated by the presence of a specific RNA opener.
Smart RNA guides are conditionally activated by the presence of a specific RNA opener.
Here we report smart RNA guides that are conditionally activated by the presence of a specific RNA opener.
Smart RNA guides are conditionally activated by the presence of a specific RNA opener.
Here we report smart RNA guides that are conditionally activated by the presence of a specific RNA opener.
Smart RNA guides are conditionally activated by the presence of a specific RNA opener.
Here we report smart RNA guides that are conditionally activated by the presence of a specific RNA opener.
Smart RNA guides are conditionally activated by the presence of a specific RNA opener.
Here we report smart RNA guides that are conditionally activated by the presence of a specific RNA opener.
The SmartGuide design allows spacer and opener to be independent, affording design flexibility.
Contrary to most previous approaches, the design affords ample freedom as spacer and the opener are independent.
The SmartGuide design allows spacer and opener to be independent, affording design flexibility.
Contrary to most previous approaches, the design affords ample freedom as spacer and the opener are independent.
The SmartGuide design allows spacer and opener to be independent, affording design flexibility.
Contrary to most previous approaches, the design affords ample freedom as spacer and the opener are independent.
The SmartGuide design allows spacer and opener to be independent, affording design flexibility.
Contrary to most previous approaches, the design affords ample freedom as spacer and the opener are independent.
The SmartGuide design allows spacer and opener to be independent, affording design flexibility.
Contrary to most previous approaches, the design affords ample freedom as spacer and the opener are independent.
The SmartGuide design allows spacer and opener to be independent, affording design flexibility.
Contrary to most previous approaches, the design affords ample freedom as spacer and the opener are independent.
The SmartGuide design allows spacer and opener to be independent, affording design flexibility.
Contrary to most previous approaches, the design affords ample freedom as spacer and the opener are independent.
In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation.
Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation.
Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation.
Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation.
Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation.
Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation.
Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation.
Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
Approval Evidence
1 source4 linked approval claimsfirst-pass slug smart-rna-guides
Here we report smart RNA guides that are conditionally activated by the presence of a specific RNA opener.
Source:
application scopesupports
SmartGuides were operated with a panel of miRNA relevant for human health and composed into Boolean logic circuits.
We demonstrate this flexibility by operating SmartGuides activated by a panel of miRNA relevant for human health, and by composing SmartGuides in Boolean logic circuits.
Source:
conditional activationsupports
Smart RNA guides are conditionally activated by the presence of a specific RNA opener.
Here we report smart RNA guides that are conditionally activated by the presence of a specific RNA opener.
Source:
design propertysupports
The SmartGuide design allows spacer and opener to be independent, affording design flexibility.
Contrary to most previous approaches, the design affords ample freedom as spacer and the opener are independent.
Source:
mammalian cell test outcomemixed
In mammalian cells, SmartGuides validated the basic tenets of the design but also revealed challenges for in vivo operation.
Lastly, we test the SmartGuides in mammalian cells - validating the basics tenets of the design, but also highlighting the challenges that remain to be lifted for in-vivo operation.
Source:
Comparisons
Source-backed strengths
A key strength is conditional activation by a specific RNA opener, which enables input-dependent control of CRISPR-Cas9. The design reportedly allows spacer and opener to be independent, and the system was applied across a panel of miRNAs relevant to human health and in Boolean logic circuits.
Compared with ribozyme-flanked gRNA
Smart RNA guides and ribozyme-flanked gRNA address a similar problem space.
Shared frame: same top-level item type
Compared with sgRNA
Smart RNA guides and sgRNA address a similar problem space.
Shared frame: same top-level item type
Compared with tet-controlled riboregulatory module
Smart RNA guides and tet-controlled riboregulatory module address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
Ranked Citations
- 1.