Source 1primary paper2022Analytical Chemistry
Toolkit/photoactivated CRISPR/Cas12a strategy
photoactivated CRISPR/Cas12a strategy
Also known as: photoactivatable CRISPR/Cas12a strategy
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The photoactivated CRISPR/Cas12a strategy is a light-gated one-pot DETECTR nucleic acid detection system. It uses a photocleavable complementary ssDNA to transiently block crRNA activity during early recombinase polymerase amplification (RPA) and activates Cas12a after brief 365 nm ultraviolet exposure for sensitive detection.
Usefulness & Problems
Why this is useful
This strategy is useful for integrating amplification and CRISPR/Cas12a readout in a single reaction while preserving high analytical sensitivity. The reported one-pot format can reduce amplicon contamination risk and lower the threshold for point-of-care molecular diagnostics.
Problem solved
A central problem in one-pot CRISPR diagnostics is that prematurely active Cas12a can interfere with target amplification before sufficient amplicon accumulates. This strategy addresses that timing conflict by temporarily suppressing crRNA function until the amplification reaction has progressed through the early exponential phase.
Problem links
Need a controllable or interpretable biological readout
DerivedThe photoactivated CRISPR/Cas12a strategy is a light-gated, multi-component one-pot DETECTR system for high-sensitivity nucleic acid detection. It uses a photocleavable complementary ssDNA to temporarily block crRNA activity and then activates Cas12a after brief 365 nm ultraviolet exposure.
Need controllable genome or transcript editing
DerivedThe photoactivated CRISPR/Cas12a strategy is a light-gated, multi-component one-pot DETECTR system for high-sensitivity nucleic acid detection. It uses a photocleavable complementary ssDNA to temporarily block crRNA activity and then activates Cas12a after brief 365 nm ultraviolet exposure.
Need precise spatiotemporal control with light input
DerivedThe photoactivated CRISPR/Cas12a strategy is a light-gated, multi-component one-pot DETECTR system for high-sensitivity nucleic acid detection. It uses a photocleavable complementary ssDNA to temporarily block crRNA activity and then activates Cas12a after brief 365 nm ultraviolet exposure.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
crrna blocking by complementary ssdnalight-triggered temporal activation of cas12aPhotocleavagePhotocleavagePhotocleavageTechniques
No technique tags yet.
Target processes
diagnosticeditingInput: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: multi component delivery burdenimplementation constraint: spectral hardware requirementoperating role: sensorswitch architecture: cleavageswitch architecture: multi component
The construct design includes a photocleavable complementary ssDNA that blocks crRNA prior to activation. The assay is implemented as a one-pot DETECTR workflow with RPA and requires brief 365 nm ultraviolet exposure to release Cas12a activity after sufficient amplicon accumulation.
The available evidence is limited to a single 2022 study and focuses on diagnostic use rather than genome editing applications. Practical performance across sample types, robustness outside the reported assay context, and any effects of 365 nm ultraviolet exposure on assay components are not described in the supplied evidence.
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
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
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 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 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.
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.
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 source3 linked approval claimsfirst-pass slug photoactivated-crispr-cas12a-strategy
This study proposes a photoactivated CRISPR/Cas12a strategy to achieve one-pot high-sensitivity nucleic acid detection.
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:
performancesupports
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.
Source:
practical advantagesupports
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.
Source:
Comparisons
Source-backed strengths
The method achieved a reported sensitivity of 2.5 copies within 40 minutes in a one-pot format. Its light-triggered activation provides temporal control over Cas12a activity and supports contamination-avoiding assay integration for molecular diagnosis.
Source:
This one-pot method achieved a sensitivity of 2.5 copies within 40 min.
Compared with NIR light-activated CRISPR-dCas9/Cas9 system
photoactivated CRISPR/Cas12a strategy and NIR light-activated CRISPR-dCas9/Cas9 system address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing; shared mechanisms: photocleavage; same primary input modality: light
Compared with photoactivatable CRISPR/Cas12a system
photoactivated CRISPR/Cas12a strategy and photoactivatable CRISPR/Cas12a system address a similar problem space because they share diagnostic, editing.
Shared frame: same top-level item type; shared target processes: diagnostic, editing; shared mechanisms: photocleavage; same primary input modality: light
Relative tradeoffs: appears more independently replicated; looks easier to implement in practice.
Compared with photoactivatable nanoCRISPR/Cas9 system
photoactivated CRISPR/Cas12a strategy and photoactivatable nanoCRISPR/Cas9 system address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing; shared mechanisms: photocleavage; same primary input modality: light
Ranked Citations
- 1.