Toolkit/photo-sensitive circular gRNAs

photo-sensitive circular gRNAs

RNA Element·Research·Since 2022

Also known as: circular gRNA, cyclically caged guide RNAs

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

Summary

Photo-sensitive circular gRNAs are cyclically caged guide RNAs that enable light-activated CRISPR/Cas9- and Cpf1-mediated genome editing. They are designed for spatiotemporal control of editing and are activated by photocleavage of the circularized guide.

Usefulness & Problems

Why this is useful

This tool is useful for controlling when and where CRISPR editing occurs using light. The cited work presents it as an improved method for precise spatiotemporal manipulation of gene editing and reports efficient editing with CRISPR/Cas9 and Cpf1.

Source:

Together, our work provides a significantly improved method to precisely manipulate where and when genes are edited.

Source:

we reported a spatiotemporal and efficient CRISPR/Cas9 and Cpf1-mediated editing with photo-sensitive circular gRNAs

Problem solved

It addresses the problem of limited temporal and spatial precision in genome editing. The reported design also reduces unwanted activity before activation, as a bow-knot-type gRNA showed no background editing without light irradiation.

Source:

Together, our work provides a significantly improved method to precisely manipulate where and when genes are edited.

Problem links

Need controllable genome or transcript editing

Derived

Photo-sensitive circular gRNAs are cyclically caged guide RNAs that enable light-activated CRISPR/Cas9- and Cpf1-mediated genome editing. They are designed for spatiotemporal control of editing and are activated by photocleavage of the circularized guide.

Need precise spatiotemporal control with light input

Derived

Photo-sensitive circular gRNAs are cyclically caged guide RNAs that enable light-activated CRISPR/Cas9- and Cpf1-mediated genome editing. They are designed for spatiotemporal control of editing and are activated by photocleavage of the circularized guide.

Taxonomy & Function

Primary hierarchy

Mechanism Branch

Component: A low-level RNA part used inside a larger architecture that realizes a mechanism.

Mechanisms

Photocleavage

Techniques

No technique tags yet.

Target processes

editing

Input: Light

Implementation Constraints

cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: spectral hardware requirementoperating role: regulatorswitch architecture: cleavage

Implementation involves circularized, photo-sensitive guide RNAs used with CRISPR/Cas9 or Cpf1 systems. The evidence supports covalent cyclization and chemical modification conceptually, but it does not provide construct architecture, delivery method, or cofactor requirements.

The supplied evidence does not specify the photocleavable chemistry, illumination wavelength, activation kinetics, or quantitative editing efficiencies. Validation is currently supported by a single cited study, with embryo editing mentioned only for MSTN.

Validation

Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication

Supporting Sources

Ranked Claims

Claim 1application scopesupports2022Source 1needs review

The method is presented as an improved way to precisely manipulate where and when genes are edited.

Together, our work provides a significantly improved method to precisely manipulate where and when genes are edited.
Claim 2application scopesupports2022Source 1needs review

The method is presented as an improved way to precisely manipulate where and when genes are edited.

Together, our work provides a significantly improved method to precisely manipulate where and when genes are edited.
Claim 3application scopesupports2022Source 1needs review

The method is presented as an improved way to precisely manipulate where and when genes are edited.

Together, our work provides a significantly improved method to precisely manipulate where and when genes are edited.
Claim 4application scopesupports2022Source 1needs review

The method is presented as an improved way to precisely manipulate where and when genes are edited.

Together, our work provides a significantly improved method to precisely manipulate where and when genes are edited.
Claim 5application scopesupports2022Source 1needs review

The method is presented as an improved way to precisely manipulate where and when genes are edited.

Together, our work provides a significantly improved method to precisely manipulate where and when genes are edited.
Claim 6application scopesupports2022Source 1needs review

The method is presented as an improved way to precisely manipulate where and when genes are edited.

Together, our work provides a significantly improved method to precisely manipulate where and when genes are edited.
Claim 7application scopesupports2022Source 1needs review

The method is presented as an improved way to precisely manipulate where and when genes are edited.

Together, our work provides a significantly improved method to precisely manipulate where and when genes are edited.
Claim 8background activitysupports2022Source 1needs review

A bow-knot-type gRNA showed no background editing in the absence of light irradiation.

a new bow-knot-type gRNA showed no background editing in the absence of light irradiation
Claim 9background activitysupports2022Source 1needs review

A bow-knot-type gRNA showed no background editing in the absence of light irradiation.

a new bow-knot-type gRNA showed no background editing in the absence of light irradiation
Claim 10background activitysupports2022Source 1needs review

A bow-knot-type gRNA showed no background editing in the absence of light irradiation.

a new bow-knot-type gRNA showed no background editing in the absence of light irradiation
Claim 11background activitysupports2022Source 1needs review

A bow-knot-type gRNA showed no background editing in the absence of light irradiation.

a new bow-knot-type gRNA showed no background editing in the absence of light irradiation
Claim 12background activitysupports2022Source 1needs review

A bow-knot-type gRNA showed no background editing in the absence of light irradiation.

a new bow-knot-type gRNA showed no background editing in the absence of light irradiation
Claim 13background activitysupports2022Source 1needs review

A bow-knot-type gRNA showed no background editing in the absence of light irradiation.

a new bow-knot-type gRNA showed no background editing in the absence of light irradiation
Claim 14background activitysupports2022Source 1needs review

A bow-knot-type gRNA showed no background editing in the absence of light irradiation.

a new bow-knot-type gRNA showed no background editing in the absence of light irradiation
Claim 15embryo editingsupports2022Source 1needs review

Light-mediated MSTN gene editing was achieved in embryos.

We have also achieved light-mediated MSTN gene editing in embryos
Claim 16embryo editingsupports2022Source 1needs review

Light-mediated MSTN gene editing was achieved in embryos.

We have also achieved light-mediated MSTN gene editing in embryos
Claim 17embryo editingsupports2022Source 1needs review

Light-mediated MSTN gene editing was achieved in embryos.

We have also achieved light-mediated MSTN gene editing in embryos
Claim 18embryo editingsupports2022Source 1needs review

Light-mediated MSTN gene editing was achieved in embryos.

We have also achieved light-mediated MSTN gene editing in embryos
Claim 19embryo editingsupports2022Source 1needs review

Light-mediated MSTN gene editing was achieved in embryos.

We have also achieved light-mediated MSTN gene editing in embryos
Claim 20embryo editingsupports2022Source 1needs review

Light-mediated MSTN gene editing was achieved in embryos.

We have also achieved light-mediated MSTN gene editing in embryos
Claim 21embryo editingsupports2022Source 1needs review

Light-mediated MSTN gene editing was achieved in embryos.

We have also achieved light-mediated MSTN gene editing in embryos
Claim 22engineering method advantagesupports2022Source 1needs review

The circular gRNA approach uses only two or three pre-installed photolabile substituents followed by simple covalent cyclization and is described as a more robust synthesis approach than heavily modified gRNAs.

This approach relies on only two or three pre-installed photolabile substituents followed by a simple covalent cyclization, which provides a robust synthesize approach in comparison to heavily modified gRNAs.
Claim 23engineering method advantagesupports2022Source 1needs review

The circular gRNA approach uses only two or three pre-installed photolabile substituents followed by simple covalent cyclization and is described as a more robust synthesis approach than heavily modified gRNAs.

This approach relies on only two or three pre-installed photolabile substituents followed by a simple covalent cyclization, which provides a robust synthesize approach in comparison to heavily modified gRNAs.
Claim 24engineering method advantagesupports2022Source 1needs review

The circular gRNA approach uses only two or three pre-installed photolabile substituents followed by simple covalent cyclization and is described as a more robust synthesis approach than heavily modified gRNAs.

This approach relies on only two or three pre-installed photolabile substituents followed by a simple covalent cyclization, which provides a robust synthesize approach in comparison to heavily modified gRNAs.
Claim 25engineering method advantagesupports2022Source 1needs review

The circular gRNA approach uses only two or three pre-installed photolabile substituents followed by simple covalent cyclization and is described as a more robust synthesis approach than heavily modified gRNAs.

This approach relies on only two or three pre-installed photolabile substituents followed by a simple covalent cyclization, which provides a robust synthesize approach in comparison to heavily modified gRNAs.
Claim 26engineering method advantagesupports2022Source 1needs review

The circular gRNA approach uses only two or three pre-installed photolabile substituents followed by simple covalent cyclization and is described as a more robust synthesis approach than heavily modified gRNAs.

This approach relies on only two or three pre-installed photolabile substituents followed by a simple covalent cyclization, which provides a robust synthesize approach in comparison to heavily modified gRNAs.
Claim 27engineering method advantagesupports2022Source 1needs review

The circular gRNA approach uses only two or three pre-installed photolabile substituents followed by simple covalent cyclization and is described as a more robust synthesis approach than heavily modified gRNAs.

This approach relies on only two or three pre-installed photolabile substituents followed by a simple covalent cyclization, which provides a robust synthesize approach in comparison to heavily modified gRNAs.
Claim 28engineering method advantagesupports2022Source 1needs review

The circular gRNA approach uses only two or three pre-installed photolabile substituents followed by simple covalent cyclization and is described as a more robust synthesis approach than heavily modified gRNAs.

This approach relies on only two or three pre-installed photolabile substituents followed by a simple covalent cyclization, which provides a robust synthesize approach in comparison to heavily modified gRNAs.
Claim 29functional capabilitysupports2022Source 1needs review

Photo-sensitive circular gRNAs enable spatiotemporal and efficient CRISPR/Cas9- and Cpf1-mediated editing.

we reported a spatiotemporal and efficient CRISPR/Cas9 and Cpf1-mediated editing with photo-sensitive circular gRNAs
Claim 30functional capabilitysupports2022Source 1needs review

Photo-sensitive circular gRNAs enable spatiotemporal and efficient CRISPR/Cas9- and Cpf1-mediated editing.

we reported a spatiotemporal and efficient CRISPR/Cas9 and Cpf1-mediated editing with photo-sensitive circular gRNAs
Claim 31functional capabilitysupports2022Source 1needs review

Photo-sensitive circular gRNAs enable spatiotemporal and efficient CRISPR/Cas9- and Cpf1-mediated editing.

we reported a spatiotemporal and efficient CRISPR/Cas9 and Cpf1-mediated editing with photo-sensitive circular gRNAs
Claim 32functional capabilitysupports2022Source 1needs review

Photo-sensitive circular gRNAs enable spatiotemporal and efficient CRISPR/Cas9- and Cpf1-mediated editing.

we reported a spatiotemporal and efficient CRISPR/Cas9 and Cpf1-mediated editing with photo-sensitive circular gRNAs
Claim 33functional capabilitysupports2022Source 1needs review

Photo-sensitive circular gRNAs enable spatiotemporal and efficient CRISPR/Cas9- and Cpf1-mediated editing.

we reported a spatiotemporal and efficient CRISPR/Cas9 and Cpf1-mediated editing with photo-sensitive circular gRNAs
Claim 34functional capabilitysupports2022Source 1needs review

Photo-sensitive circular gRNAs enable spatiotemporal and efficient CRISPR/Cas9- and Cpf1-mediated editing.

we reported a spatiotemporal and efficient CRISPR/Cas9 and Cpf1-mediated editing with photo-sensitive circular gRNAs
Claim 35functional capabilitysupports2022Source 1needs review

Photo-sensitive circular gRNAs enable spatiotemporal and efficient CRISPR/Cas9- and Cpf1-mediated editing.

we reported a spatiotemporal and efficient CRISPR/Cas9 and Cpf1-mediated editing with photo-sensitive circular gRNAs
Claim 36light gated editingsupports2022Source 1needs review

In established cells stably expressing Cas9, circular gRNA together with light irradiation directs precise cleavage of GFP and VEGFA within a pre-defined cutting region.

In established cells stably expressing Cas9, the circular gRNA in coordination with light irradiation could direct a precise cleavage of GFP and VEGFA within a pre-defined cutting region.
Claim 37light gated editingsupports2022Source 1needs review

In established cells stably expressing Cas9, circular gRNA together with light irradiation directs precise cleavage of GFP and VEGFA within a pre-defined cutting region.

In established cells stably expressing Cas9, the circular gRNA in coordination with light irradiation could direct a precise cleavage of GFP and VEGFA within a pre-defined cutting region.
Claim 38light gated editingsupports2022Source 1needs review

In established cells stably expressing Cas9, circular gRNA together with light irradiation directs precise cleavage of GFP and VEGFA within a pre-defined cutting region.

In established cells stably expressing Cas9, the circular gRNA in coordination with light irradiation could direct a precise cleavage of GFP and VEGFA within a pre-defined cutting region.
Claim 39light gated editingsupports2022Source 1needs review

In established cells stably expressing Cas9, circular gRNA together with light irradiation directs precise cleavage of GFP and VEGFA within a pre-defined cutting region.

In established cells stably expressing Cas9, the circular gRNA in coordination with light irradiation could direct a precise cleavage of GFP and VEGFA within a pre-defined cutting region.
Claim 40light gated editingsupports2022Source 1needs review

In established cells stably expressing Cas9, circular gRNA together with light irradiation directs precise cleavage of GFP and VEGFA within a pre-defined cutting region.

In established cells stably expressing Cas9, the circular gRNA in coordination with light irradiation could direct a precise cleavage of GFP and VEGFA within a pre-defined cutting region.
Claim 41light gated editingsupports2022Source 1needs review

In established cells stably expressing Cas9, circular gRNA together with light irradiation directs precise cleavage of GFP and VEGFA within a pre-defined cutting region.

In established cells stably expressing Cas9, the circular gRNA in coordination with light irradiation could direct a precise cleavage of GFP and VEGFA within a pre-defined cutting region.
Claim 42light gated editingsupports2022Source 1needs review

In established cells stably expressing Cas9, circular gRNA together with light irradiation directs precise cleavage of GFP and VEGFA within a pre-defined cutting region.

In established cells stably expressing Cas9, the circular gRNA in coordination with light irradiation could direct a precise cleavage of GFP and VEGFA within a pre-defined cutting region.

Approval Evidence

1 source5 linked approval claimsfirst-pass slug photo-sensitive-circular-grnas
we reported a spatiotemporal and efficient CRISPR/Cas9 and Cpf1-mediated editing with photo-sensitive circular gRNAs

Source:

application scopesupports

The method is presented as an improved way to precisely manipulate where and when genes are edited.

Together, our work provides a significantly improved method to precisely manipulate where and when genes are edited.

Source:

embryo editingsupports

Light-mediated MSTN gene editing was achieved in embryos.

We have also achieved light-mediated MSTN gene editing in embryos

Source:

engineering method advantagesupports

The circular gRNA approach uses only two or three pre-installed photolabile substituents followed by simple covalent cyclization and is described as a more robust synthesis approach than heavily modified gRNAs.

This approach relies on only two or three pre-installed photolabile substituents followed by a simple covalent cyclization, which provides a robust synthesize approach in comparison to heavily modified gRNAs.

Source:

functional capabilitysupports

Photo-sensitive circular gRNAs enable spatiotemporal and efficient CRISPR/Cas9- and Cpf1-mediated editing.

we reported a spatiotemporal and efficient CRISPR/Cas9 and Cpf1-mediated editing with photo-sensitive circular gRNAs

Source:

light gated editingsupports

In established cells stably expressing Cas9, circular gRNA together with light irradiation directs precise cleavage of GFP and VEGFA within a pre-defined cutting region.

In established cells stably expressing Cas9, the circular gRNA in coordination with light irradiation could direct a precise cleavage of GFP and VEGFA within a pre-defined cutting region.

Source:

Comparisons

Source-backed strengths

Reported strengths include spatiotemporal and efficient CRISPR/Cas9- and Cpf1-mediated editing. The source also states that a bow-knot-type gRNA had no detectable background editing in the absence of light and that light-mediated MSTN gene editing was achieved in embryos.

Source:

This approach relies on only two or three pre-installed photolabile substituents followed by a simple covalent cyclization, which provides a robust synthesize approach in comparison to heavily modified gRNAs.

photo-sensitive circular gRNAs and auxiliary photocleavable oligodeoxyribonucleotides complementary to 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 caged guide RNA

photo-sensitive circular gRNAs 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

photo-sensitive circular gRNAs 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

Ranked Citations

  1. 1.

    Extracted from this source document.