Toolkit/photoactivatable CRISPR/Cas9 system

photoactivatable CRISPR/Cas9 system

Multi-Component Switch·Research·Since 2021

Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.

Summary

The photoactivatable CRISPR/Cas9 system is a light-gated genome-editing configuration comprising Cas9, either a synthetic 102-nt sgRNA or a crRNA/tracrRNA pair, and blocking photocleavable oligodeoxyribonucleotides. UV irradiation in the presence of the photomodified blocking oligodeoxyribonucleotides enables photoactivatable gene editing in vitro.

Usefulness & Problems

Why this is useful

This system is useful for imposing optical control over CRISPR/Cas9-mediated editing by keeping the guide RNA functionally blocked until UV exposure. The available evidence supports in vitro light-triggered activation, but does not further quantify performance or application scope.

Source:

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.

Problem solved

It addresses the problem of making CRISPR/Cas9 editing conditional on an external light input rather than constitutively active once Cas9 and guide RNA are present. The reported design uses blocking photocleavable oligodeoxyribonucleotides to gate editing until UV irradiation.

Problem links

Need controllable genome or transcript editing

Derived

The photoactivatable CRISPR/Cas9 system is a light-gated genome-editing configuration composed of Cas9, either a synthetic 102-nt sgRNA or a crRNA/tracrRNA pair, and blocking photocleavable oligodeoxyribonucleotides. UV irradiation in the presence of the photomodified blocking oligodeoxyribonucleotides enables photoactivatable gene editing in vitro.

Need precise spatiotemporal control with light input

Derived

The photoactivatable CRISPR/Cas9 system is a light-gated genome-editing configuration composed of Cas9, either a synthetic 102-nt sgRNA or a crRNA/tracrRNA pair, and blocking photocleavable oligodeoxyribonucleotides. UV irradiation in the presence of the photomodified blocking oligodeoxyribonucleotides enables photoactivatable gene editing in vitro.

Taxonomy & Function

Primary hierarchy

Mechanism Branch

Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.

Target processes

editing

Input: 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: regulatorswitch architecture: cleavageswitch architecture: multi component

The reported components are Cas9 protein, either a synthetic 102-nt sgRNA or a guide crRNA/tracrRNA pair, and blocking photocleavable oligodeoxyribonucleotides. Activation requires UV irradiation, but the supplied evidence does not specify wavelength, dose, delivery method, or construct stoichiometry.

The evidence provided is limited to design description and an in vitro functional claim under UV irradiation. No quantitative editing efficiencies, target loci, cell-based validation, organismal use, off-target analysis, or independent replication are described in the supplied evidence.

Validation

Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication

Supporting Sources

Ranked Claims

Claim 1designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 2designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 3designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 4designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 5designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 6designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 7designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 8designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 9designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 10designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 11designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 12designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 13designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 14designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 15designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 16designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 17designsupports2021Source 1needs review

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.
Claim 18functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 19functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 20functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 21functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 22functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 23functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 24functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 25functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 26functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 27functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 28functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 29functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 30functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 31functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 32functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 33functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.
Claim 34functional effectsupports2021Source 1needs review

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.

Approval Evidence

1 source2 linked approval claimsfirst-pass slug photoactivatable-crispr-cas9-system
A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.

Source:

designsupports

A photoactivatable CRISPR/Cas9 system composed of Cas9, guide RNA, and blocking photocleavable oligodeoxyribonucleotides was designed.

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed.

Source:

functional effectsupports

UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can produce photoactivatable gene editing in vitro.

It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.

Source:

Comparisons

Source-backed strengths

The system was explicitly designed as a multi-component photoactivatable CRISPR/Cas9 configuration and supports either a single-guide RNA format or a crRNA/tracrRNA format. Source claims state that UV irradiation can induce gene editing in vitro in the presence of the photomodified blocking oligodeoxyribonucleotides.

photoactivatable CRISPR/Cas9 system 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

photoactivatable CRISPR/Cas9 system 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

photoactivatable CRISPR/Cas9 system and photoactivated CRISPR/Cas12a strategy 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. 1.
    StructuralSource 1Russian Journal of Bioorganic Chemistry2021Claim 12Claim 11Claim 11

    Extracted from this source document.