Source 1primary paper2023Chemical Science
Toolkit/methylated guide RNA for CRISPR-Cas12a
methylated guide RNA for CRISPR-Cas12a
Also known as: epigenetically modified guide RNA, m6A- or m1A-methylated gRNA
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Methylated guide RNA for CRISPR-Cas12a is a chemically modified crRNA bearing m6A or m1A marks that suppresses Cas12a activity. The methylated guide inhibits both cis- and trans-DNA cleavage, and activity can be reactivated through guide RNA demethylation.
Usefulness & Problems
Why this is useful
This tool provides a reversible RNA-level method to regulate CRISPR-Cas12a function without changing the Cas12a protein itself. Reported applications include regulation of gene expression, demethylase imaging in living cells, and controllable gene editing.
Source:
This strategy has been used in the regulation of gene expression, demethylase imaging in living cells and controllable gene editing.
Source:
The results demonstrate that the methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
Problem solved
It addresses the problem of how to switch Cas12a DNA targeting and cleavage off and back on in a controllable manner. The approach uses guide RNA methylation to deactivate Cas12a and demethylation to restore function.
Source:
This strategy has been used in the regulation of gene expression, demethylase imaging in living cells and controllable gene editing.
Problem links
Need conditional recombination or state switching
DerivedMethylated guide RNA for CRISPR-Cas12a is a chemically modified crRNA bearing m6A or m1A marks that suppresses Cas12a activity. The methylated guide inhibits both cis- and trans-DNA cleavage, and activity can be reactivated through guide RNA demethylation.
Need controllable genome or transcript editing
DerivedMethylated guide RNA for CRISPR-Cas12a is a chemically modified crRNA bearing m6A or m1A marks that suppresses Cas12a activity. The methylated guide inhibits both cis- and trans-DNA cleavage, and activity can be reactivated through guide RNA demethylation.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level RNA part used inside a larger architecture that realizes a mechanism.
Mechanisms
destabilization of guide rna secondary and tertiary structureepitranscriptomic inhibition by guide rna methylationPhotocleavageprevention of cas12a-guide rna complex assemblyreversible reactivation by demethylationTechniques
No technique tags yet.
Target processes
editingrecombinationImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: regulatorswitch architecture: cleavage
Implementation requires chemically methylated crRNA containing m6A or m1A modifications and a demethylation step for reactivation. The supplied evidence supports use in living cells, but it does not specify the demethylase identity, construct architecture, delivery format, or sequence-design constraints.
The evidence provided comes from a single 2023 Chemical Science study, so independent replication is not established here. Practical performance details such as modification-site rules, quantitative dynamic range, compatibility across Cas12a orthologs, and delivery constraints are not described in the supplied evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
m6A and m1A methylation of guide RNA inhibits both cis- and trans-DNA cleavage activities of CRISPR-Cas12a.
epigenetic modifications of guide RNA, such as m6A and m1A methylation, can effectively inhibit both the cis- and trans-DNA cleavage activities of CRISPR-Cas12a
m6A and m1A methylation of guide RNA inhibits both cis- and trans-DNA cleavage activities of CRISPR-Cas12a.
epigenetic modifications of guide RNA, such as m6A and m1A methylation, can effectively inhibit both the cis- and trans-DNA cleavage activities of CRISPR-Cas12a
m6A and m1A methylation of guide RNA inhibits both cis- and trans-DNA cleavage activities of CRISPR-Cas12a.
epigenetic modifications of guide RNA, such as m6A and m1A methylation, can effectively inhibit both the cis- and trans-DNA cleavage activities of CRISPR-Cas12a
m6A and m1A methylation of guide RNA inhibits both cis- and trans-DNA cleavage activities of CRISPR-Cas12a.
epigenetic modifications of guide RNA, such as m6A and m1A methylation, can effectively inhibit both the cis- and trans-DNA cleavage activities of CRISPR-Cas12a
m6A and m1A methylation of guide RNA inhibits both cis- and trans-DNA cleavage activities of CRISPR-Cas12a.
epigenetic modifications of guide RNA, such as m6A and m1A methylation, can effectively inhibit both the cis- and trans-DNA cleavage activities of CRISPR-Cas12a
m6A and m1A methylation of guide RNA inhibits both cis- and trans-DNA cleavage activities of CRISPR-Cas12a.
epigenetic modifications of guide RNA, such as m6A and m1A methylation, can effectively inhibit both the cis- and trans-DNA cleavage activities of CRISPR-Cas12a
m6A and m1A methylation of guide RNA inhibits both cis- and trans-DNA cleavage activities of CRISPR-Cas12a.
epigenetic modifications of guide RNA, such as m6A and m1A methylation, can effectively inhibit both the cis- and trans-DNA cleavage activities of CRISPR-Cas12a
The methylation-deactivated and demethylase-activated strategy was used for regulation of gene expression, demethylase imaging in living cells, and controllable gene editing.
This strategy has been used in the regulation of gene expression, demethylase imaging in living cells and controllable gene editing.
The methylation-deactivated and demethylase-activated strategy was used for regulation of gene expression, demethylase imaging in living cells, and controllable gene editing.
This strategy has been used in the regulation of gene expression, demethylase imaging in living cells and controllable gene editing.
The methylation-deactivated and demethylase-activated strategy was used for regulation of gene expression, demethylase imaging in living cells, and controllable gene editing.
This strategy has been used in the regulation of gene expression, demethylase imaging in living cells and controllable gene editing.
The methylation-deactivated and demethylase-activated strategy was used for regulation of gene expression, demethylase imaging in living cells, and controllable gene editing.
This strategy has been used in the regulation of gene expression, demethylase imaging in living cells and controllable gene editing.
The methylation-deactivated and demethylase-activated strategy was used for regulation of gene expression, demethylase imaging in living cells, and controllable gene editing.
This strategy has been used in the regulation of gene expression, demethylase imaging in living cells and controllable gene editing.
The methylation-deactivated and demethylase-activated strategy was used for regulation of gene expression, demethylase imaging in living cells, and controllable gene editing.
This strategy has been used in the regulation of gene expression, demethylase imaging in living cells and controllable gene editing.
The methylation-deactivated and demethylase-activated strategy was used for regulation of gene expression, demethylase imaging in living cells, and controllable gene editing.
This strategy has been used in the regulation of gene expression, demethylase imaging in living cells and controllable gene editing.
Guide RNA methylation destabilizes gRNA secondary and tertiary structure, preventing Cas12a-gRNA complex assembly and decreasing DNA targeting ability.
The underlying mechanism is that methylations destabilize the secondary and tertiary structure of gRNA which prevents the assembly of the Cas12a-gRNA nuclease complex, leading to decreased DNA targeting ability.
Guide RNA methylation destabilizes gRNA secondary and tertiary structure, preventing Cas12a-gRNA complex assembly and decreasing DNA targeting ability.
The underlying mechanism is that methylations destabilize the secondary and tertiary structure of gRNA which prevents the assembly of the Cas12a-gRNA nuclease complex, leading to decreased DNA targeting ability.
Guide RNA methylation destabilizes gRNA secondary and tertiary structure, preventing Cas12a-gRNA complex assembly and decreasing DNA targeting ability.
The underlying mechanism is that methylations destabilize the secondary and tertiary structure of gRNA which prevents the assembly of the Cas12a-gRNA nuclease complex, leading to decreased DNA targeting ability.
Guide RNA methylation destabilizes gRNA secondary and tertiary structure, preventing Cas12a-gRNA complex assembly and decreasing DNA targeting ability.
The underlying mechanism is that methylations destabilize the secondary and tertiary structure of gRNA which prevents the assembly of the Cas12a-gRNA nuclease complex, leading to decreased DNA targeting ability.
Guide RNA methylation destabilizes gRNA secondary and tertiary structure, preventing Cas12a-gRNA complex assembly and decreasing DNA targeting ability.
The underlying mechanism is that methylations destabilize the secondary and tertiary structure of gRNA which prevents the assembly of the Cas12a-gRNA nuclease complex, leading to decreased DNA targeting ability.
Guide RNA methylation destabilizes gRNA secondary and tertiary structure, preventing Cas12a-gRNA complex assembly and decreasing DNA targeting ability.
The underlying mechanism is that methylations destabilize the secondary and tertiary structure of gRNA which prevents the assembly of the Cas12a-gRNA nuclease complex, leading to decreased DNA targeting ability.
Guide RNA methylation destabilizes gRNA secondary and tertiary structure, preventing Cas12a-gRNA complex assembly and decreasing DNA targeting ability.
The underlying mechanism is that methylations destabilize the secondary and tertiary structure of gRNA which prevents the assembly of the Cas12a-gRNA nuclease complex, leading to decreased DNA targeting ability.
The inhibitory effects of guide RNA methylation on CRISPR-Cas12a are reversible through demethylation of gRNA by demethylases.
We also demonstrate that these effects are reversible through the demethylation of gRNA by demethylases.
The inhibitory effects of guide RNA methylation on CRISPR-Cas12a are reversible through demethylation of gRNA by demethylases.
We also demonstrate that these effects are reversible through the demethylation of gRNA by demethylases.
The inhibitory effects of guide RNA methylation on CRISPR-Cas12a are reversible through demethylation of gRNA by demethylases.
We also demonstrate that these effects are reversible through the demethylation of gRNA by demethylases.
The inhibitory effects of guide RNA methylation on CRISPR-Cas12a are reversible through demethylation of gRNA by demethylases.
We also demonstrate that these effects are reversible through the demethylation of gRNA by demethylases.
The inhibitory effects of guide RNA methylation on CRISPR-Cas12a are reversible through demethylation of gRNA by demethylases.
We also demonstrate that these effects are reversible through the demethylation of gRNA by demethylases.
The inhibitory effects of guide RNA methylation on CRISPR-Cas12a are reversible through demethylation of gRNA by demethylases.
We also demonstrate that these effects are reversible through the demethylation of gRNA by demethylases.
The inhibitory effects of guide RNA methylation on CRISPR-Cas12a are reversible through demethylation of gRNA by demethylases.
We also demonstrate that these effects are reversible through the demethylation of gRNA by demethylases.
A minimum of three adenine methylated nucleotides is required to completely inhibit Cas12a nuclease activity.
A minimum of three adenine methylated nucleotides are required to completely inhibit the nuclease activity.
minimum adenine methylated nucleotides for complete inhibition 3 nucleotides
A minimum of three adenine methylated nucleotides is required to completely inhibit Cas12a nuclease activity.
A minimum of three adenine methylated nucleotides are required to completely inhibit the nuclease activity.
minimum adenine methylated nucleotides for complete inhibition 3 nucleotides
A minimum of three adenine methylated nucleotides is required to completely inhibit Cas12a nuclease activity.
A minimum of three adenine methylated nucleotides are required to completely inhibit the nuclease activity.
minimum adenine methylated nucleotides for complete inhibition 3 nucleotides
A minimum of three adenine methylated nucleotides is required to completely inhibit Cas12a nuclease activity.
A minimum of three adenine methylated nucleotides are required to completely inhibit the nuclease activity.
minimum adenine methylated nucleotides for complete inhibition 3 nucleotides
A minimum of three adenine methylated nucleotides is required to completely inhibit Cas12a nuclease activity.
A minimum of three adenine methylated nucleotides are required to completely inhibit the nuclease activity.
minimum adenine methylated nucleotides for complete inhibition 3 nucleotides
A minimum of three adenine methylated nucleotides is required to completely inhibit Cas12a nuclease activity.
A minimum of three adenine methylated nucleotides are required to completely inhibit the nuclease activity.
minimum adenine methylated nucleotides for complete inhibition 3 nucleotides
A minimum of three adenine methylated nucleotides is required to completely inhibit Cas12a nuclease activity.
A minimum of three adenine methylated nucleotides are required to completely inhibit the nuclease activity.
minimum adenine methylated nucleotides for complete inhibition 3 nucleotides
The methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The results demonstrate that the methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The results demonstrate that the methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The results demonstrate that the methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The results demonstrate that the methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The results demonstrate that the methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The results demonstrate that the methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
The results demonstrate that the methylation-deactivated and demethylase-activated strategy is a promising tool for regulation of the CRISPR-Cas12a system.
Approval Evidence
1 source4 linked approval claimsfirst-pass slug methylated-guide-rna-for-crispr-cas12a
epigenetic modifications of guide RNA, such as m6A and m1A methylation, can effectively inhibit both the cis- and trans-DNA cleavage activities of CRISPR-Cas12a
Source:
activity regulationsupports
m6A and m1A methylation of guide RNA inhibits both cis- and trans-DNA cleavage activities of CRISPR-Cas12a.
epigenetic modifications of guide RNA, such as m6A and m1A methylation, can effectively inhibit both the cis- and trans-DNA cleavage activities of CRISPR-Cas12a
Source:
mechanismsupports
Guide RNA methylation destabilizes gRNA secondary and tertiary structure, preventing Cas12a-gRNA complex assembly and decreasing DNA targeting ability.
The underlying mechanism is that methylations destabilize the secondary and tertiary structure of gRNA which prevents the assembly of the Cas12a-gRNA nuclease complex, leading to decreased DNA targeting ability.
Source:
reversibilitysupports
The inhibitory effects of guide RNA methylation on CRISPR-Cas12a are reversible through demethylation of gRNA by demethylases.
We also demonstrate that these effects are reversible through the demethylation of gRNA by demethylases.
Source:
threshold requirementsupports
A minimum of three adenine methylated nucleotides is required to completely inhibit Cas12a nuclease activity.
A minimum of three adenine methylated nucleotides are required to completely inhibit the nuclease activity.
Source:
Comparisons
Source-backed strengths
m6A and m1A modification of the guide RNA was reported to effectively inhibit both cis- and trans-DNA cleavage activities of CRISPR-Cas12a. The study also links this functional suppression to a defined structural mechanism in which methylation destabilizes guide RNA secondary and tertiary structure and prevents Cas12a-gRNA complex assembly.
Compared with antisense oligonucleotides
methylated guide RNA for CRISPR-Cas12a and antisense oligonucleotides address a similar problem space because they share editing, recombination.
Shared frame: same top-level item type; shared target processes: editing, recombination
Strengths here: looks easier to implement in practice.
Compared with light-controlled crRNA
methylated guide RNA for CRISPR-Cas12a 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
Strengths here: looks easier to implement in practice.
Compared with photo-sensitive circular gRNAs
methylated guide RNA for CRISPR-Cas12a 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
Strengths here: looks easier to implement in practice.
Ranked Citations
- 1.