Toolkit/methylated guide RNA for CRISPR-Cas12a

methylated guide RNA for CRISPR-Cas12a

RNA Element·Research·Since 2023

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

Derived

Methylated guide RNA for CRISPR-Cas12a is a chemically modified crRNA in which m6A or m1A marks are installed to suppress Cas12a function. These epitranscriptomic modifications inhibit both cis- and trans-DNA cleavage, and activity can be restored by guide RNA demethylation.

Need controllable genome or transcript editing

Derived

Methylated guide RNA for CRISPR-Cas12a is a chemically modified crRNA in which m6A or m1A marks are installed to suppress Cas12a function. These epitranscriptomic modifications inhibit both cis- and trans-DNA cleavage, and activity can be restored by guide RNA demethylation.

Taxonomy & Function

Implementation 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

Claim 1activity regulationsupports2023Source 1needs review

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
Claim 2activity regulationsupports2023Source 1needs review

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
Claim 3activity regulationsupports2023Source 1needs review

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
Claim 4activity regulationsupports2023Source 1needs review

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
Claim 5activity regulationsupports2023Source 1needs review

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
Claim 6activity regulationsupports2023Source 1needs review

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
Claim 7activity regulationsupports2023Source 1needs review

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
Claim 8activity regulationsupports2023Source 1needs review

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
Claim 9activity regulationsupports2023Source 1needs review

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
Claim 10activity regulationsupports2023Source 1needs review

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
Claim 11activity regulationsupports2023Source 1needs review

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
Claim 12activity regulationsupports2023Source 1needs review

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
Claim 13activity regulationsupports2023Source 1needs review

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
Claim 14activity regulationsupports2023Source 1needs review

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
Claim 15activity regulationsupports2023Source 1needs review

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
Claim 16activity regulationsupports2023Source 1needs review

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
Claim 17activity regulationsupports2023Source 1needs review

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
Claim 18application scopesupports2023Source 1needs review

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.
Claim 19application scopesupports2023Source 1needs review

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.
Claim 20application scopesupports2023Source 1needs review

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.
Claim 21application scopesupports2023Source 1needs review

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.
Claim 22application scopesupports2023Source 1needs review

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.
Claim 23application scopesupports2023Source 1needs review

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.
Claim 24application scopesupports2023Source 1needs review

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.
Claim 25application scopesupports2023Source 1needs review

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.
Claim 26application scopesupports2023Source 1needs review

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.
Claim 27application scopesupports2023Source 1needs review

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.
Claim 28mechanismsupports2023Source 1needs review

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.
Claim 29mechanismsupports2023Source 1needs review

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.
Claim 30mechanismsupports2023Source 1needs review

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.
Claim 31mechanismsupports2023Source 1needs review

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.
Claim 32mechanismsupports2023Source 1needs review

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.
Claim 33mechanismsupports2023Source 1needs review

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.
Claim 34mechanismsupports2023Source 1needs review

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.
Claim 35mechanismsupports2023Source 1needs review

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.
Claim 36mechanismsupports2023Source 1needs review

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.
Claim 37mechanismsupports2023Source 1needs review

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.
Claim 38mechanismsupports2023Source 1needs review

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.
Claim 39mechanismsupports2023Source 1needs review

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.
Claim 40mechanismsupports2023Source 1needs review

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.
Claim 41mechanismsupports2023Source 1needs review

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.
Claim 42mechanismsupports2023Source 1needs review

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.
Claim 43mechanismsupports2023Source 1needs review

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.
Claim 44mechanismsupports2023Source 1needs review

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.
Claim 45reversibilitysupports2023Source 1needs review

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.
Claim 46reversibilitysupports2023Source 1needs review

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.
Claim 47reversibilitysupports2023Source 1needs review

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.
Claim 48reversibilitysupports2023Source 1needs review

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.
Claim 49reversibilitysupports2023Source 1needs review

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.
Claim 50reversibilitysupports2023Source 1needs review

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.
Claim 51reversibilitysupports2023Source 1needs review

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.
Claim 52reversibilitysupports2023Source 1needs review

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.
Claim 53reversibilitysupports2023Source 1needs review

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.
Claim 54reversibilitysupports2023Source 1needs review

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.
Claim 55reversibilitysupports2023Source 1needs review

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.
Claim 56reversibilitysupports2023Source 1needs review

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.
Claim 57reversibilitysupports2023Source 1needs review

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.
Claim 58reversibilitysupports2023Source 1needs review

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.
Claim 59reversibilitysupports2023Source 1needs review

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.
Claim 60reversibilitysupports2023Source 1needs review

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.
Claim 61reversibilitysupports2023Source 1needs review

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.
Claim 62threshold requirementsupports2023Source 1needs review

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
Claim 63threshold requirementsupports2023Source 1needs review

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
Claim 64threshold requirementsupports2023Source 1needs review

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
Claim 65threshold requirementsupports2023Source 1needs review

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
Claim 66threshold requirementsupports2023Source 1needs review

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
Claim 67threshold requirementsupports2023Source 1needs review

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
Claim 68threshold requirementsupports2023Source 1needs review

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
Claim 69threshold requirementsupports2023Source 1needs review

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
Claim 70threshold requirementsupports2023Source 1needs review

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
Claim 71threshold requirementsupports2023Source 1needs review

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
Claim 72threshold requirementsupports2023Source 1needs review

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
Claim 73threshold requirementsupports2023Source 1needs review

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
Claim 74threshold requirementsupports2023Source 1needs review

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
Claim 75threshold requirementsupports2023Source 1needs review

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
Claim 76threshold requirementsupports2023Source 1needs review

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
Claim 77threshold requirementsupports2023Source 1needs review

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
Claim 78threshold requirementsupports2023Source 1needs review

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
Claim 79tool promisesupports2023Source 1needs review

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.
Claim 80tool promisesupports2023Source 1needs review

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.
Claim 81tool promisesupports2023Source 1needs review

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.
Claim 82tool promisesupports2023Source 1needs review

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.
Claim 83tool promisesupports2023Source 1needs review

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.
Claim 84tool promisesupports2023Source 1needs review

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.
Claim 85tool promisesupports2023Source 1needs review

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.
Claim 86tool promisesupports2023Source 1needs review

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.
Claim 87tool promisesupports2023Source 1needs review

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.
Claim 88tool promisesupports2023Source 1needs review

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.

methylated guide RNA for CRISPR-Cas12a and photoactivatable CRISPR/Cas12a system address a similar problem space because they share editing, recombination.

Shared frame: shared target processes: editing, recombination; shared mechanisms: photocleavage

Strengths here: looks easier to implement in practice.

Relative tradeoffs: appears more independently replicated.

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.

methylated guide RNA for CRISPR-Cas12a and synthetically engineered guide RNA address a similar problem space because they share editing, recombination.

Shared frame: same top-level item type; shared target processes: editing, recombination

Ranked Citations

  1. 1.
    StructuralSource 1Chemical Science2023Claim 1Claim 12Claim 12

    Extracted from this source document.