Source 1primary paper2021International Journal of Molecular Sciences
Toolkit/auxiliary photocleavable oligodeoxyribonucleotides complementary to crRNA
auxiliary photocleavable oligodeoxyribonucleotides complementary to crRNA
Also known as: PC-DNAs
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Auxiliary photocleavable oligodeoxyribonucleotides complementary to crRNA (PC-DNAs) are inhibitory oligonucleotide components of a photoactivatable nanoCRISPR/Cas9 system. They hybridize to crRNA to suppress Cas9 function before illumination and are photocleaved by 365 nm UV light to release crRNA and restore gene-editing activity.
Usefulness & Problems
Why this is useful
PC-DNAs provide light-gated control over CRISPR/Cas9 activity, enabling temporal activation of gene editing by external irradiation. In the reported nanoCRISPR/Cas9 format, this strategy was used to create a large functional difference between pre-irradiation and post-irradiation Cas9 activity.
Problem solved
This tool addresses the problem of keeping crRNA-dependent Cas9 editing inactive until a defined light stimulus is applied. It specifically solves reversible crRNA blocking in a nanoparticle-associated CRISPR/Cas9 system using photocleavable complementary oligodeoxyribonucleotides.
Problem links
Need controllable genome or transcript editing
DerivedAuxiliary photocleavable oligodeoxyribonucleotides complementary to crRNA (PC-DNAs) are inhibitory oligonucleotide components of a photoactivatable nanoCRISPR/Cas9 system. They hybridize to crRNA to suppress Cas9 function before illumination and are photocleaved by 365 nm UV light to release crRNA and restore gene-editing activity.
Need precise spatiotemporal control with light input
DerivedAuxiliary photocleavable oligodeoxyribonucleotides complementary to crRNA (PC-DNAs) are inhibitory oligonucleotide components of a photoactivatable nanoCRISPR/Cas9 system. They hybridize to crRNA to suppress Cas9 function before illumination and are photocleaved by 365 nm UV light to release crRNA and restore gene-editing activity.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level RNA part used inside a larger architecture that realizes a mechanism.
Techniques
No technique tags yet.
Target processes
editingInput: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: spectral hardware requirementoperating role: regulatorswitch architecture: cleavage
PC-DNAs were designed as oligodeoxyribonucleotides complementary to crRNA and used in a system where the photocleavable oligonucleotides were immobilized on carbon nanoparticles. Practical parameters reported as important included oligonucleotide length, number of photocleavable linkers, UV irradiation time, and nanoparticle type; 365 nm irradiation was used for activation.
The evidence provided is limited to a single 2021 study describing a nanoCRISPR/Cas9 implementation. Validation details beyond light-dependent Cas9 activity control, including broader organismal testing, editing outcomes across targets, and independent replication, are not supplied here.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
The authors proposed a photoactivatable CRISPR/Cas9 gene-editing system based on photocleavable oligodeoxyribonucleotides complementary to crRNA.
Here, we proposed a new approach to engineering a photoactivatable CRISPR/Cas9 gene-editing system. The novel nanoCRISPR/Cas9 system is based on the use of auxiliary photocleavable oligodeoxyribonucleotides (PC-DNAs) complementary to crRNA.
The authors proposed a photoactivatable CRISPR/Cas9 gene-editing system based on photocleavable oligodeoxyribonucleotides complementary to crRNA.
Here, we proposed a new approach to engineering a photoactivatable CRISPR/Cas9 gene-editing system. The novel nanoCRISPR/Cas9 system is based on the use of auxiliary photocleavable oligodeoxyribonucleotides (PC-DNAs) complementary to crRNA.
The authors proposed a photoactivatable CRISPR/Cas9 gene-editing system based on photocleavable oligodeoxyribonucleotides complementary to crRNA.
Here, we proposed a new approach to engineering a photoactivatable CRISPR/Cas9 gene-editing system. The novel nanoCRISPR/Cas9 system is based on the use of auxiliary photocleavable oligodeoxyribonucleotides (PC-DNAs) complementary to crRNA.
The authors proposed a photoactivatable CRISPR/Cas9 gene-editing system based on photocleavable oligodeoxyribonucleotides complementary to crRNA.
Here, we proposed a new approach to engineering a photoactivatable CRISPR/Cas9 gene-editing system. The novel nanoCRISPR/Cas9 system is based on the use of auxiliary photocleavable oligodeoxyribonucleotides (PC-DNAs) complementary to crRNA.
The authors proposed a photoactivatable CRISPR/Cas9 gene-editing system based on photocleavable oligodeoxyribonucleotides complementary to crRNA.
Here, we proposed a new approach to engineering a photoactivatable CRISPR/Cas9 gene-editing system. The novel nanoCRISPR/Cas9 system is based on the use of auxiliary photocleavable oligodeoxyribonucleotides (PC-DNAs) complementary to crRNA.
The authors proposed a photoactivatable CRISPR/Cas9 gene-editing system based on photocleavable oligodeoxyribonucleotides complementary to crRNA.
Here, we proposed a new approach to engineering a photoactivatable CRISPR/Cas9 gene-editing system. The novel nanoCRISPR/Cas9 system is based on the use of auxiliary photocleavable oligodeoxyribonucleotides (PC-DNAs) complementary to crRNA.
The authors proposed a photoactivatable CRISPR/Cas9 gene-editing system based on photocleavable oligodeoxyribonucleotides complementary to crRNA.
Here, we proposed a new approach to engineering a photoactivatable CRISPR/Cas9 gene-editing system. The novel nanoCRISPR/Cas9 system is based on the use of auxiliary photocleavable oligodeoxyribonucleotides (PC-DNAs) complementary to crRNA.
Immobilizing photocleavable oligonucleotides on carbon nanoparticles blocks crRNA and corresponding Cas9 activity before UV irradiation, and UV irradiation at 365 nm releases crRNA and restores Cas9 activity.
Immobilizing PC-DNAs on the surface of carbon nanoparticles through 3'-terminal pyrene residue provided sufficient blocking of crRNA (and corresponding Cas9 activity) before UV irradiation and allows for crRNA release after UV irradiation at 365 nm, which restores Cas9 activity.
UV irradiation wavelength 365 nm
Immobilizing photocleavable oligonucleotides on carbon nanoparticles blocks crRNA and corresponding Cas9 activity before UV irradiation, and UV irradiation at 365 nm releases crRNA and restores Cas9 activity.
Immobilizing PC-DNAs on the surface of carbon nanoparticles through 3'-terminal pyrene residue provided sufficient blocking of crRNA (and corresponding Cas9 activity) before UV irradiation and allows for crRNA release after UV irradiation at 365 nm, which restores Cas9 activity.
UV irradiation wavelength 365 nm
Immobilizing photocleavable oligonucleotides on carbon nanoparticles blocks crRNA and corresponding Cas9 activity before UV irradiation, and UV irradiation at 365 nm releases crRNA and restores Cas9 activity.
Immobilizing PC-DNAs on the surface of carbon nanoparticles through 3'-terminal pyrene residue provided sufficient blocking of crRNA (and corresponding Cas9 activity) before UV irradiation and allows for crRNA release after UV irradiation at 365 nm, which restores Cas9 activity.
UV irradiation wavelength 365 nm
Immobilizing photocleavable oligonucleotides on carbon nanoparticles blocks crRNA and corresponding Cas9 activity before UV irradiation, and UV irradiation at 365 nm releases crRNA and restores Cas9 activity.
Immobilizing PC-DNAs on the surface of carbon nanoparticles through 3'-terminal pyrene residue provided sufficient blocking of crRNA (and corresponding Cas9 activity) before UV irradiation and allows for crRNA release after UV irradiation at 365 nm, which restores Cas9 activity.
UV irradiation wavelength 365 nm
Immobilizing photocleavable oligonucleotides on carbon nanoparticles blocks crRNA and corresponding Cas9 activity before UV irradiation, and UV irradiation at 365 nm releases crRNA and restores Cas9 activity.
Immobilizing PC-DNAs on the surface of carbon nanoparticles through 3'-terminal pyrene residue provided sufficient blocking of crRNA (and corresponding Cas9 activity) before UV irradiation and allows for crRNA release after UV irradiation at 365 nm, which restores Cas9 activity.
UV irradiation wavelength 365 nm
Immobilizing photocleavable oligonucleotides on carbon nanoparticles blocks crRNA and corresponding Cas9 activity before UV irradiation, and UV irradiation at 365 nm releases crRNA and restores Cas9 activity.
Immobilizing PC-DNAs on the surface of carbon nanoparticles through 3'-terminal pyrene residue provided sufficient blocking of crRNA (and corresponding Cas9 activity) before UV irradiation and allows for crRNA release after UV irradiation at 365 nm, which restores Cas9 activity.
UV irradiation wavelength 365 nm
Immobilizing photocleavable oligonucleotides on carbon nanoparticles blocks crRNA and corresponding Cas9 activity before UV irradiation, and UV irradiation at 365 nm releases crRNA and restores Cas9 activity.
Immobilizing PC-DNAs on the surface of carbon nanoparticles through 3'-terminal pyrene residue provided sufficient blocking of crRNA (and corresponding Cas9 activity) before UV irradiation and allows for crRNA release after UV irradiation at 365 nm, which restores Cas9 activity.
UV irradiation wavelength 365 nm
The authors optimized blocking oligonucleotide length, linker number, irradiation time, and carbon nanoparticle type, and identified the carbon-encapsulated iron nanoparticle version as the most promising because it gave the greatest before-versus-after irradiation functional activity difference.
We optimized the length of blocking photocleavable oligonucleotide, number of linkers, time of irradiation, and the type of carbon nanoparticles. Based on the results, we consider the nanoCRISPR/Cas9 system involving carbon-encapsulated iron nanoparticles the most promising. It provides the greatest difference of functional activity before/after irradiation
The authors optimized blocking oligonucleotide length, linker number, irradiation time, and carbon nanoparticle type, and identified the carbon-encapsulated iron nanoparticle version as the most promising because it gave the greatest before-versus-after irradiation functional activity difference.
We optimized the length of blocking photocleavable oligonucleotide, number of linkers, time of irradiation, and the type of carbon nanoparticles. Based on the results, we consider the nanoCRISPR/Cas9 system involving carbon-encapsulated iron nanoparticles the most promising. It provides the greatest difference of functional activity before/after irradiation
The authors optimized blocking oligonucleotide length, linker number, irradiation time, and carbon nanoparticle type, and identified the carbon-encapsulated iron nanoparticle version as the most promising because it gave the greatest before-versus-after irradiation functional activity difference.
We optimized the length of blocking photocleavable oligonucleotide, number of linkers, time of irradiation, and the type of carbon nanoparticles. Based on the results, we consider the nanoCRISPR/Cas9 system involving carbon-encapsulated iron nanoparticles the most promising. It provides the greatest difference of functional activity before/after irradiation
The authors optimized blocking oligonucleotide length, linker number, irradiation time, and carbon nanoparticle type, and identified the carbon-encapsulated iron nanoparticle version as the most promising because it gave the greatest before-versus-after irradiation functional activity difference.
We optimized the length of blocking photocleavable oligonucleotide, number of linkers, time of irradiation, and the type of carbon nanoparticles. Based on the results, we consider the nanoCRISPR/Cas9 system involving carbon-encapsulated iron nanoparticles the most promising. It provides the greatest difference of functional activity before/after irradiation
The authors optimized blocking oligonucleotide length, linker number, irradiation time, and carbon nanoparticle type, and identified the carbon-encapsulated iron nanoparticle version as the most promising because it gave the greatest before-versus-after irradiation functional activity difference.
We optimized the length of blocking photocleavable oligonucleotide, number of linkers, time of irradiation, and the type of carbon nanoparticles. Based on the results, we consider the nanoCRISPR/Cas9 system involving carbon-encapsulated iron nanoparticles the most promising. It provides the greatest difference of functional activity before/after irradiation
The authors optimized blocking oligonucleotide length, linker number, irradiation time, and carbon nanoparticle type, and identified the carbon-encapsulated iron nanoparticle version as the most promising because it gave the greatest before-versus-after irradiation functional activity difference.
We optimized the length of blocking photocleavable oligonucleotide, number of linkers, time of irradiation, and the type of carbon nanoparticles. Based on the results, we consider the nanoCRISPR/Cas9 system involving carbon-encapsulated iron nanoparticles the most promising. It provides the greatest difference of functional activity before/after irradiation
The authors optimized blocking oligonucleotide length, linker number, irradiation time, and carbon nanoparticle type, and identified the carbon-encapsulated iron nanoparticle version as the most promising because it gave the greatest before-versus-after irradiation functional activity difference.
We optimized the length of blocking photocleavable oligonucleotide, number of linkers, time of irradiation, and the type of carbon nanoparticles. Based on the results, we consider the nanoCRISPR/Cas9 system involving carbon-encapsulated iron nanoparticles the most promising. It provides the greatest difference of functional activity before/after irradiation
The carbon-encapsulated iron nanoparticle nanoCRISPR/Cas9 system could prospectively support magnetic field-controlled delivery and UV-induced spatiotemporal gene editing.
and can be used in prospective for magnetic field-controlled delivery of CRISPR system into the target cells or tissues and spatiotemporal gene editing induced by UV irradiation.
The carbon-encapsulated iron nanoparticle nanoCRISPR/Cas9 system could prospectively support magnetic field-controlled delivery and UV-induced spatiotemporal gene editing.
and can be used in prospective for magnetic field-controlled delivery of CRISPR system into the target cells or tissues and spatiotemporal gene editing induced by UV irradiation.
The carbon-encapsulated iron nanoparticle nanoCRISPR/Cas9 system could prospectively support magnetic field-controlled delivery and UV-induced spatiotemporal gene editing.
and can be used in prospective for magnetic field-controlled delivery of CRISPR system into the target cells or tissues and spatiotemporal gene editing induced by UV irradiation.
The carbon-encapsulated iron nanoparticle nanoCRISPR/Cas9 system could prospectively support magnetic field-controlled delivery and UV-induced spatiotemporal gene editing.
and can be used in prospective for magnetic field-controlled delivery of CRISPR system into the target cells or tissues and spatiotemporal gene editing induced by UV irradiation.
The carbon-encapsulated iron nanoparticle nanoCRISPR/Cas9 system could prospectively support magnetic field-controlled delivery and UV-induced spatiotemporal gene editing.
and can be used in prospective for magnetic field-controlled delivery of CRISPR system into the target cells or tissues and spatiotemporal gene editing induced by UV irradiation.
The carbon-encapsulated iron nanoparticle nanoCRISPR/Cas9 system could prospectively support magnetic field-controlled delivery and UV-induced spatiotemporal gene editing.
and can be used in prospective for magnetic field-controlled delivery of CRISPR system into the target cells or tissues and spatiotemporal gene editing induced by UV irradiation.
The carbon-encapsulated iron nanoparticle nanoCRISPR/Cas9 system could prospectively support magnetic field-controlled delivery and UV-induced spatiotemporal gene editing.
and can be used in prospective for magnetic field-controlled delivery of CRISPR system into the target cells or tissues and spatiotemporal gene editing induced by UV irradiation.
Approval Evidence
1 source2 linked approval claimsfirst-pass slug auxiliary-photocleavable-oligodeoxyribonucleotides-complementary-to-crrna
The novel nanoCRISPR/Cas9 system is based on the use of auxiliary photocleavable oligodeoxyribonucleotides (PC-DNAs) complementary to crRNA.
Source:
engineering approachsupports
The authors proposed a photoactivatable CRISPR/Cas9 gene-editing system based on photocleavable oligodeoxyribonucleotides complementary to crRNA.
Here, we proposed a new approach to engineering a photoactivatable CRISPR/Cas9 gene-editing system. The novel nanoCRISPR/Cas9 system is based on the use of auxiliary photocleavable oligodeoxyribonucleotides (PC-DNAs) complementary to crRNA.
Source:
mechanism of controlsupports
Immobilizing photocleavable oligonucleotides on carbon nanoparticles blocks crRNA and corresponding Cas9 activity before UV irradiation, and UV irradiation at 365 nm releases crRNA and restores Cas9 activity.
Immobilizing PC-DNAs on the surface of carbon nanoparticles through 3'-terminal pyrene residue provided sufficient blocking of crRNA (and corresponding Cas9 activity) before UV irradiation and allows for crRNA release after UV irradiation at 365 nm, which restores Cas9 activity.
Source:
Comparisons
Source-backed strengths
The system was experimentally optimized across blocking oligonucleotide length, photocleavable linker number, irradiation time, and carbon nanoparticle type. Among the tested formulations, the carbon-encapsulated iron nanoparticle version produced the greatest difference in functional activity before versus after irradiation.
Source:
Here, we proposed a new approach to engineering a photoactivatable CRISPR/Cas9 gene-editing system. The novel nanoCRISPR/Cas9 system is based on the use of auxiliary photocleavable oligodeoxyribonucleotides (PC-DNAs) complementary to crRNA.
Source:
We optimized the length of blocking photocleavable oligonucleotide, number of linkers, time of irradiation, and the type of carbon nanoparticles. Based on the results, we consider the nanoCRISPR/Cas9 system involving carbon-encapsulated iron nanoparticles the most promising. It provides the greatest difference of functional activity before/after irradiation
Compared with caged guide RNA
auxiliary photocleavable oligodeoxyribonucleotides complementary to crRNA and caged guide RNA 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 light-controlled crRNA
auxiliary photocleavable oligodeoxyribonucleotides complementary to crRNA and light-controlled crRNA 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 photo-sensitive circular gRNAs
auxiliary photocleavable oligodeoxyribonucleotides complementary to crRNA 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; shared mechanisms: photocleavage; same primary input modality: light
Ranked Citations
- 1.