Source 1primary paper2024Angewandte Chemie
Toolkit/CRISPR/Cas9-inducible DNAzyme probe
CRISPR/Cas9-inducible DNAzyme probe
Also known as: CRISPR-DNAzyme
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
CRISPR-DNAzyme is a three-stranded DNAzyme probe engineered for in situ imaging of nuclear Zn2+ in living cells. Its catalytic activity is initially blocked by a 20-bp CRISPR/Cas9 recognition site and is activated in the nucleus after Cas9/sgRNA cleavage forms the active DNAzyme structure.
Usefulness & Problems
Why this is useful
This tool enables nuclear Zn2+ imaging with CRISPR/Cas9-dependent activation, allowing signal generation to be restricted to a defined intracellular context. Integration with photoactivation and a Boolean logic gate was reported to provide superior spatiotemporal control for dynamic monitoring of nuclear Zn2+ in HeLa cells and mice.
Source:
With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
Problem solved
It addresses the problem of achieving in situ, nucleus-specific imaging of Zn2+ while suppressing DNAzyme activity until a programmed activation event occurs. The design specifically couples Zn2+ sensing to CRISPR/Cas9 cleavage so that the catalytic probe is assembled only after target-site processing in the nucleus.
Source:
With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete method used to build, optimize, or evolve an engineered system.
Mechanisms
boolean logic gatingconditional dnazyme activationcrispr/cas9-mediated cleavagephotoactivationPhotocleavageTechniques
Computational DesignTarget processes
editinglocalizationInput: Light
Implementation Constraints
The probe is described as a three-stranded DNAzyme construct containing a 20-bp CRISPR/Cas9 recognition site that blocks catalysis before cleavage. Activation requires Cas9 and sgRNA, and the reported enhanced control involved integration with a photoactivation strategy and Boolean logic gate; the supplied evidence does not specify construct sequences, delivery format, or illumination parameters.
The supplied evidence is limited to a single 2024 source and focuses on nuclear Zn2+ imaging rather than broader applications. Quantitative performance metrics, sequence-level design details, and evidence for use beyond HeLa cells and mice are not provided in the supplied material.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Observations
successMammalian Cell Lineapplication demo
Inferred from claim c2 during normalization. The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells. Derived from claim c2. Quoted text: With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
Source:
successMammalian Cell Lineapplication demo
Inferred from claim c2 during normalization. The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells. Derived from claim c2. Quoted text: With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
Source:
successMammalian Cell Lineapplication demo
Inferred from claim c2 during normalization. The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells. Derived from claim c2. Quoted text: With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
Source:
successMammalian Cell Lineapplication demo
Inferred from claim c2 during normalization. The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells. Derived from claim c2. Quoted text: With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
Source:
successMammalian Cell Lineapplication demo
Inferred from claim c2 during normalization. The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells. Derived from claim c2. Quoted text: With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
Source:
successMammalian Cell Lineapplication demo
Inferred from claim c2 during normalization. The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells. Derived from claim c2. Quoted text: With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
Source:
successMammalian Cell Lineapplication demo
Inferred from claim c2 during normalization. The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells. Derived from claim c2. Quoted text: With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
Source:
Supporting Sources
Ranked Claims
The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells.
With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells.
With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells.
With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells.
With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells.
With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells.
With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells.
With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
Integrating the CRISPR-DNAzyme system with photoactivation strategy and Boolean logic gate provided superior spatiotemporal control imaging for dynamic monitoring of nuclear Zn2+ in HeLa cells and mice.
Moreover, the superiority of CRISPR‐DNAzyme for spatiotemporal control imaging was demonstrated by integrating it with photoactivation strategy and Boolean logic gate for dynamic monitoring nuclear Zn 2+ in both HeLa cells and mice.
Integrating the CRISPR-DNAzyme system with photoactivation strategy and Boolean logic gate provided superior spatiotemporal control imaging for dynamic monitoring of nuclear Zn2+ in HeLa cells and mice.
Moreover, the superiority of CRISPR‐DNAzyme for spatiotemporal control imaging was demonstrated by integrating it with photoactivation strategy and Boolean logic gate for dynamic monitoring nuclear Zn 2+ in both HeLa cells and mice.
Integrating the CRISPR-DNAzyme system with photoactivation strategy and Boolean logic gate provided superior spatiotemporal control imaging for dynamic monitoring of nuclear Zn2+ in HeLa cells and mice.
Moreover, the superiority of CRISPR‐DNAzyme for spatiotemporal control imaging was demonstrated by integrating it with photoactivation strategy and Boolean logic gate for dynamic monitoring nuclear Zn 2+ in both HeLa cells and mice.
Integrating the CRISPR-DNAzyme system with photoactivation strategy and Boolean logic gate provided superior spatiotemporal control imaging for dynamic monitoring of nuclear Zn2+ in HeLa cells and mice.
Moreover, the superiority of CRISPR‐DNAzyme for spatiotemporal control imaging was demonstrated by integrating it with photoactivation strategy and Boolean logic gate for dynamic monitoring nuclear Zn 2+ in both HeLa cells and mice.
Integrating the CRISPR-DNAzyme system with photoactivation strategy and Boolean logic gate provided superior spatiotemporal control imaging for dynamic monitoring of nuclear Zn2+ in HeLa cells and mice.
Moreover, the superiority of CRISPR‐DNAzyme for spatiotemporal control imaging was demonstrated by integrating it with photoactivation strategy and Boolean logic gate for dynamic monitoring nuclear Zn 2+ in both HeLa cells and mice.
Integrating the CRISPR-DNAzyme system with photoactivation strategy and Boolean logic gate provided superior spatiotemporal control imaging for dynamic monitoring of nuclear Zn2+ in HeLa cells and mice.
Moreover, the superiority of CRISPR‐DNAzyme for spatiotemporal control imaging was demonstrated by integrating it with photoactivation strategy and Boolean logic gate for dynamic monitoring nuclear Zn 2+ in both HeLa cells and mice.
Integrating the CRISPR-DNAzyme system with photoactivation strategy and Boolean logic gate provided superior spatiotemporal control imaging for dynamic monitoring of nuclear Zn2+ in HeLa cells and mice.
Moreover, the superiority of CRISPR‐DNAzyme for spatiotemporal control imaging was demonstrated by integrating it with photoactivation strategy and Boolean logic gate for dynamic monitoring nuclear Zn 2+ in both HeLa cells and mice.
The three-stranded DNAzyme probe contains a 20-bp CRISPR/Cas9 recognition site that blocks DNAzyme activity until Cas9/sgRNA cleavage forms the catalytic DNAzyme structure in the nucleus.
We developed a three‐stranded DNAzyme probe (TSDP) that contained a 20‐base‐pair (20‐bp) recognition site of a CRISPR/Cas9, which blocks the DNAzyme activity. When Cas9, with its specialized nuclear localization function, forms an active complex with sgRNA within the cell nucleus, it cleaves the TSDP at the recognition site, resulting in the in situ formation of catalytic DNAzyme structure.
recognition site length 20 bp
The three-stranded DNAzyme probe contains a 20-bp CRISPR/Cas9 recognition site that blocks DNAzyme activity until Cas9/sgRNA cleavage forms the catalytic DNAzyme structure in the nucleus.
We developed a three‐stranded DNAzyme probe (TSDP) that contained a 20‐base‐pair (20‐bp) recognition site of a CRISPR/Cas9, which blocks the DNAzyme activity. When Cas9, with its specialized nuclear localization function, forms an active complex with sgRNA within the cell nucleus, it cleaves the TSDP at the recognition site, resulting in the in situ formation of catalytic DNAzyme structure.
recognition site length 20 bp
The three-stranded DNAzyme probe contains a 20-bp CRISPR/Cas9 recognition site that blocks DNAzyme activity until Cas9/sgRNA cleavage forms the catalytic DNAzyme structure in the nucleus.
We developed a three‐stranded DNAzyme probe (TSDP) that contained a 20‐base‐pair (20‐bp) recognition site of a CRISPR/Cas9, which blocks the DNAzyme activity. When Cas9, with its specialized nuclear localization function, forms an active complex with sgRNA within the cell nucleus, it cleaves the TSDP at the recognition site, resulting in the in situ formation of catalytic DNAzyme structure.
recognition site length 20 bp
The three-stranded DNAzyme probe contains a 20-bp CRISPR/Cas9 recognition site that blocks DNAzyme activity until Cas9/sgRNA cleavage forms the catalytic DNAzyme structure in the nucleus.
We developed a three‐stranded DNAzyme probe (TSDP) that contained a 20‐base‐pair (20‐bp) recognition site of a CRISPR/Cas9, which blocks the DNAzyme activity. When Cas9, with its specialized nuclear localization function, forms an active complex with sgRNA within the cell nucleus, it cleaves the TSDP at the recognition site, resulting in the in situ formation of catalytic DNAzyme structure.
recognition site length 20 bp
The three-stranded DNAzyme probe contains a 20-bp CRISPR/Cas9 recognition site that blocks DNAzyme activity until Cas9/sgRNA cleavage forms the catalytic DNAzyme structure in the nucleus.
We developed a three‐stranded DNAzyme probe (TSDP) that contained a 20‐base‐pair (20‐bp) recognition site of a CRISPR/Cas9, which blocks the DNAzyme activity. When Cas9, with its specialized nuclear localization function, forms an active complex with sgRNA within the cell nucleus, it cleaves the TSDP at the recognition site, resulting in the in situ formation of catalytic DNAzyme structure.
recognition site length 20 bp
The three-stranded DNAzyme probe contains a 20-bp CRISPR/Cas9 recognition site that blocks DNAzyme activity until Cas9/sgRNA cleavage forms the catalytic DNAzyme structure in the nucleus.
We developed a three‐stranded DNAzyme probe (TSDP) that contained a 20‐base‐pair (20‐bp) recognition site of a CRISPR/Cas9, which blocks the DNAzyme activity. When Cas9, with its specialized nuclear localization function, forms an active complex with sgRNA within the cell nucleus, it cleaves the TSDP at the recognition site, resulting in the in situ formation of catalytic DNAzyme structure.
recognition site length 20 bp
The three-stranded DNAzyme probe contains a 20-bp CRISPR/Cas9 recognition site that blocks DNAzyme activity until Cas9/sgRNA cleavage forms the catalytic DNAzyme structure in the nucleus.
We developed a three‐stranded DNAzyme probe (TSDP) that contained a 20‐base‐pair (20‐bp) recognition site of a CRISPR/Cas9, which blocks the DNAzyme activity. When Cas9, with its specialized nuclear localization function, forms an active complex with sgRNA within the cell nucleus, it cleaves the TSDP at the recognition site, resulting in the in situ formation of catalytic DNAzyme structure.
recognition site length 20 bp
This conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and provides new analytical methods for nuclear metal-associated biology.
Collectively, this conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and thus provides new analytical methods for nuclear metal‐associated biology.
This conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and provides new analytical methods for nuclear metal-associated biology.
Collectively, this conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and thus provides new analytical methods for nuclear metal‐associated biology.
This conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and provides new analytical methods for nuclear metal-associated biology.
Collectively, this conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and thus provides new analytical methods for nuclear metal‐associated biology.
This conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and provides new analytical methods for nuclear metal-associated biology.
Collectively, this conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and thus provides new analytical methods for nuclear metal‐associated biology.
This conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and provides new analytical methods for nuclear metal-associated biology.
Collectively, this conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and thus provides new analytical methods for nuclear metal‐associated biology.
This conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and provides new analytical methods for nuclear metal-associated biology.
Collectively, this conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and thus provides new analytical methods for nuclear metal‐associated biology.
This conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and provides new analytical methods for nuclear metal-associated biology.
Collectively, this conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and thus provides new analytical methods for nuclear metal‐associated biology.
Approval Evidence
1 source4 linked approval claimsfirst-pass slug crispr-cas9-inducible-dnazyme-probe
With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells. Moreover, the superiority of CRISPR‐DNAzyme for spatiotemporal control imaging was demonstrated by integrating it with photoactivation strategy and Boolean logic gate
Source:
application capabilitysupports
The CRISPR/Cas9-inducible DNAzyme design enables in situ imaging of nuclear Zn2+ in living cells.
With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells.
Source:
comparative performancesupports
Integrating the CRISPR-DNAzyme system with photoactivation strategy and Boolean logic gate provided superior spatiotemporal control imaging for dynamic monitoring of nuclear Zn2+ in HeLa cells and mice.
Moreover, the superiority of CRISPR‐DNAzyme for spatiotemporal control imaging was demonstrated by integrating it with photoactivation strategy and Boolean logic gate for dynamic monitoring nuclear Zn 2+ in both HeLa cells and mice.
Source:
design mechanismsupports
The three-stranded DNAzyme probe contains a 20-bp CRISPR/Cas9 recognition site that blocks DNAzyme activity until Cas9/sgRNA cleavage forms the catalytic DNAzyme structure in the nucleus.
We developed a three‐stranded DNAzyme probe (TSDP) that contained a 20‐base‐pair (20‐bp) recognition site of a CRISPR/Cas9, which blocks the DNAzyme activity. When Cas9, with its specialized nuclear localization function, forms an active complex with sgRNA within the cell nucleus, it cleaves the TSDP at the recognition site, resulting in the in situ formation of catalytic DNAzyme structure.
Source:
toolbox expansionsupports
This conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and provides new analytical methods for nuclear metal-associated biology.
Collectively, this conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and thus provides new analytical methods for nuclear metal‐associated biology.
Source:
Comparisons
Source-backed strengths
The reported system was demonstrated for imaging nuclear Zn2+ in living cells. Source literature further states that combining the CRISPR-DNAzyme with photoactivation and Boolean logic gating improved spatiotemporal control for dynamic nuclear Zn2+ imaging in HeLa cells and mice.
Source:
Moreover, the superiority of CRISPR‐DNAzyme for spatiotemporal control imaging was demonstrated by integrating it with photoactivation strategy and Boolean logic gate for dynamic monitoring nuclear Zn 2+ in both HeLa cells and mice.
Ranked Citations
- 1.