Source 1primary paper2022Analytical Chemistry
Toolkit/photocleavable complementary ssDNA-blocked crRNA
photocleavable complementary ssDNA-blocked crRNA
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Photocleavable complementary ssDNA-blocked crRNA is a light-gated CRISPR/Cas12a construct design in which a photocleavable complementary single-stranded DNA transiently blocks crRNA function. In a one-pot DETECTR workflow, brief 365 nm ultraviolet exposure removes this block after recombinase polymerase amplification has progressed, thereby activating Cas12a at a controlled time.
Usefulness & Problems
Why this is useful
This construct is useful for temporally separating nucleic acid amplification from Cas12a activity in a single reaction vessel. The reported one-pot photoactivated CRISPR/Cas12a format also reduces amplicon contamination risk and is described as lowering the threshold for point-of-care molecular diagnostics.
Problem solved
It addresses the incompatibility between early-phase RPA and premature Cas12a activation in one-pot DETECTR assays. By keeping crRNA inactive until light exposure, the system allows sufficient amplicon accumulation before CRISPR-based detection begins.
Problem links
Need precise spatiotemporal control with light input
DerivedPhotocleavable complementary ssDNA-blocked crRNA is a light-gated CRISPR/Cas12a construct design in which a photocleavable complementary single-stranded DNA transiently blocks crRNA function. In a one-pot DETECTR workflow, brief 365 nm ultraviolet exposure removes this block after recombinase polymerase amplification has progressed, thereby activating Cas12a at a controlled time.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Mechanisms
light-triggered temporal gating of crispr/cas12a activationnucleic acid hybridization blockingPhotocleavageTechniques
No technique tags yet.
Target processes
No target processes tagged yet.
Input: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: spectral hardware requirementoperating role: sensorswitch architecture: cleavage
Implementation involves hybridizing a photocleavable complementary ssDNA to the crRNA to suppress Cas12a activity until illumination. The reported workflow is a one-pot DETECTR assay coupled to RPA, with activation triggered by brief 365 nm ultraviolet exposure after early exponential amplification.
The evidence is limited to a single 2022 study in Analytical Chemistry and does not provide broader cross-platform or independent validation. The available evidence also does not specify sequence design rules, photocleavable linker chemistry, or performance across multiple targets, sample types, or organisms.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Photocleavable complementary ssDNA blocks crRNA so that RPA amplification can proceed through the early exponential phase without interference from activated Cas12a, and Cas12a can then be activated by brief 365 nm ultraviolet exposure after sufficient amplicon accumulation.
Using photocleavable complementary ssDNA to block crRNA, RPA amplification can smoothly pass through the exponential interval without being affected by activated Cas12a in the critical early stage. After enough amplicons were produced, the Cas12a test was activated by short bursts of ultraviolet radiation at 365 nm.
ultraviolet activation wavelength 365 nm
Photocleavable complementary ssDNA blocks crRNA so that RPA amplification can proceed through the early exponential phase without interference from activated Cas12a, and Cas12a can then be activated by brief 365 nm ultraviolet exposure after sufficient amplicon accumulation.
Using photocleavable complementary ssDNA to block crRNA, RPA amplification can smoothly pass through the exponential interval without being affected by activated Cas12a in the critical early stage. After enough amplicons were produced, the Cas12a test was activated by short bursts of ultraviolet radiation at 365 nm.
ultraviolet activation wavelength 365 nm
Photocleavable complementary ssDNA blocks crRNA so that RPA amplification can proceed through the early exponential phase without interference from activated Cas12a, and Cas12a can then be activated by brief 365 nm ultraviolet exposure after sufficient amplicon accumulation.
Using photocleavable complementary ssDNA to block crRNA, RPA amplification can smoothly pass through the exponential interval without being affected by activated Cas12a in the critical early stage. After enough amplicons were produced, the Cas12a test was activated by short bursts of ultraviolet radiation at 365 nm.
ultraviolet activation wavelength 365 nm
Photocleavable complementary ssDNA blocks crRNA so that RPA amplification can proceed through the early exponential phase without interference from activated Cas12a, and Cas12a can then be activated by brief 365 nm ultraviolet exposure after sufficient amplicon accumulation.
Using photocleavable complementary ssDNA to block crRNA, RPA amplification can smoothly pass through the exponential interval without being affected by activated Cas12a in the critical early stage. After enough amplicons were produced, the Cas12a test was activated by short bursts of ultraviolet radiation at 365 nm.
ultraviolet activation wavelength 365 nm
Photocleavable complementary ssDNA blocks crRNA so that RPA amplification can proceed through the early exponential phase without interference from activated Cas12a, and Cas12a can then be activated by brief 365 nm ultraviolet exposure after sufficient amplicon accumulation.
Using photocleavable complementary ssDNA to block crRNA, RPA amplification can smoothly pass through the exponential interval without being affected by activated Cas12a in the critical early stage. After enough amplicons were produced, the Cas12a test was activated by short bursts of ultraviolet radiation at 365 nm.
ultraviolet activation wavelength 365 nm
Photocleavable complementary ssDNA blocks crRNA so that RPA amplification can proceed through the early exponential phase without interference from activated Cas12a, and Cas12a can then be activated by brief 365 nm ultraviolet exposure after sufficient amplicon accumulation.
Using photocleavable complementary ssDNA to block crRNA, RPA amplification can smoothly pass through the exponential interval without being affected by activated Cas12a in the critical early stage. After enough amplicons were produced, the Cas12a test was activated by short bursts of ultraviolet radiation at 365 nm.
ultraviolet activation wavelength 365 nm
Photocleavable complementary ssDNA blocks crRNA so that RPA amplification can proceed through the early exponential phase without interference from activated Cas12a, and Cas12a can then be activated by brief 365 nm ultraviolet exposure after sufficient amplicon accumulation.
Using photocleavable complementary ssDNA to block crRNA, RPA amplification can smoothly pass through the exponential interval without being affected by activated Cas12a in the critical early stage. After enough amplicons were produced, the Cas12a test was activated by short bursts of ultraviolet radiation at 365 nm.
ultraviolet activation wavelength 365 nm
The one-pot photoactivated CRISPR/Cas12a method achieved a sensitivity of 2.5 copies within 40 minutes.
This one-pot method achieved a sensitivity of 2.5 copies within 40 min.
sensitivity 2.5 copiestime to result 40 min
The one-pot photoactivated CRISPR/Cas12a method achieved a sensitivity of 2.5 copies within 40 minutes.
This one-pot method achieved a sensitivity of 2.5 copies within 40 min.
sensitivity 2.5 copiestime to result 40 min
The one-pot photoactivated CRISPR/Cas12a method achieved a sensitivity of 2.5 copies within 40 minutes.
This one-pot method achieved a sensitivity of 2.5 copies within 40 min.
sensitivity 2.5 copiestime to result 40 min
The one-pot photoactivated CRISPR/Cas12a method achieved a sensitivity of 2.5 copies within 40 minutes.
This one-pot method achieved a sensitivity of 2.5 copies within 40 min.
sensitivity 2.5 copiestime to result 40 min
The one-pot photoactivated CRISPR/Cas12a method achieved a sensitivity of 2.5 copies within 40 minutes.
This one-pot method achieved a sensitivity of 2.5 copies within 40 min.
sensitivity 2.5 copiestime to result 40 min
The one-pot photoactivated CRISPR/Cas12a method achieved a sensitivity of 2.5 copies within 40 minutes.
This one-pot method achieved a sensitivity of 2.5 copies within 40 min.
sensitivity 2.5 copiestime to result 40 min
The one-pot photoactivated CRISPR/Cas12a method achieved a sensitivity of 2.5 copies within 40 minutes.
This one-pot method achieved a sensitivity of 2.5 copies within 40 min.
sensitivity 2.5 copiestime to result 40 min
The one-pot photoactivated CRISPR/Cas12a method can effectively avoid amplicon contamination and lower the threshold for point-of-care molecular diagnostics.
This simple and sensitive one-pot method can effectively avoid amplicon contamination and lower the threshold for molecular diagnostics in POC.
The one-pot photoactivated CRISPR/Cas12a method can effectively avoid amplicon contamination and lower the threshold for point-of-care molecular diagnostics.
This simple and sensitive one-pot method can effectively avoid amplicon contamination and lower the threshold for molecular diagnostics in POC.
The one-pot photoactivated CRISPR/Cas12a method can effectively avoid amplicon contamination and lower the threshold for point-of-care molecular diagnostics.
This simple and sensitive one-pot method can effectively avoid amplicon contamination and lower the threshold for molecular diagnostics in POC.
The one-pot photoactivated CRISPR/Cas12a method can effectively avoid amplicon contamination and lower the threshold for point-of-care molecular diagnostics.
This simple and sensitive one-pot method can effectively avoid amplicon contamination and lower the threshold for molecular diagnostics in POC.
The one-pot photoactivated CRISPR/Cas12a method can effectively avoid amplicon contamination and lower the threshold for point-of-care molecular diagnostics.
This simple and sensitive one-pot method can effectively avoid amplicon contamination and lower the threshold for molecular diagnostics in POC.
The one-pot photoactivated CRISPR/Cas12a method can effectively avoid amplicon contamination and lower the threshold for point-of-care molecular diagnostics.
This simple and sensitive one-pot method can effectively avoid amplicon contamination and lower the threshold for molecular diagnostics in POC.
The one-pot photoactivated CRISPR/Cas12a method can effectively avoid amplicon contamination and lower the threshold for point-of-care molecular diagnostics.
This simple and sensitive one-pot method can effectively avoid amplicon contamination and lower the threshold for molecular diagnostics in POC.
Approval Evidence
1 source1 linked approval claimfirst-pass slug photocleavable-complementary-ssdna-blocked-crrna
Using photocleavable complementary ssDNA to block crRNA
Source:
mechanismsupports
Photocleavable complementary ssDNA blocks crRNA so that RPA amplification can proceed through the early exponential phase without interference from activated Cas12a, and Cas12a can then be activated by brief 365 nm ultraviolet exposure after sufficient amplicon accumulation.
Using photocleavable complementary ssDNA to block crRNA, RPA amplification can smoothly pass through the exponential interval without being affected by activated Cas12a in the critical early stage. After enough amplicons were produced, the Cas12a test was activated by short bursts of ultraviolet radiation at 365 nm.
Source:
Comparisons
Source-backed strengths
The source reports that the one-pot photoactivated CRISPR/Cas12a method achieved a sensitivity of 2.5 copies within 40 minutes. It also provides externally triggered temporal control of Cas12a activation using brief 365 nm ultraviolet illumination.
Source:
This one-pot method achieved a sensitivity of 2.5 copies within 40 min.
Compared with alkynyl-functionalized photocleavable linker
photocleavable complementary ssDNA-blocked crRNA and alkynyl-functionalized photocleavable linker address a similar problem space.
Shared frame: same top-level item type; shared mechanisms: photocleavage; same primary input modality: light
Compared with Opto-Casp8-V1
photocleavable complementary ssDNA-blocked crRNA and Opto-Casp8-V1 address a similar problem space.
Shared frame: same top-level item type; shared mechanisms: photocleavage; same primary input modality: light
Compared with Opto-Casp8-V2
photocleavable complementary ssDNA-blocked crRNA and Opto-Casp8-V2 address a similar problem space.
Shared frame: same top-level item type; shared mechanisms: photocleavage; same primary input modality: light
Ranked Citations
- 1.