Source 1review2016Annual Review of Biochemistry
Toolkit/guide RNA
guide RNA
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Guide RNA is the programmable RNA component of the CRISPR/Cas9 system that directs Cas9 to sequence-defined genomic sites. Altering the guide RNA sequence retargets Cas9 to new DNA loci for sequence-specific genome editing.
Usefulness & Problems
Why this is useful
Guide RNA makes Cas9 readily reprogrammable, enabling sequence-specific genome engineering in diverse organisms by changing an RNA sequence rather than re-engineering the protein. The cited literature describes Cas9-based editing as several magnitudes easier to develop than prior sequence-specific gene-editing approaches, and guide RNA modification provides a route to modulate specificity.
Source:
With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics.
Problem solved
Guide RNA addresses the problem of how to redirect a DNA endonuclease to arbitrary genomic targets without redesigning the nuclease itself. It also helps address the need to tune CRISPR/Cas9 specificity through guide RNA modifications.
Source:
With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics.
Problem links
Need controllable genome or transcript editing
DerivedGuide RNA is the programmable RNA component of the CRISPR/Cas9 system that directs Cas9 to sequence-defined genomic sites. Altering the guide RNA sequence retargets Cas9 to new DNA loci for sequence-specific genome editing.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level RNA part used inside a larger architecture that realizes a mechanism.
Mechanisms
dna endonuclease-mediated genome editingrna-guided dna targetingsequence-programmed target recognitionTechniques
No technique tags yet.
Target processes
editingImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: regulator
Implementation requires pairing the guide RNA with Cas9, because the evidence describes guide RNA as the element that programs Cas9 target choice. Practical construct details, delivery formats, and specific guide RNA chemistries are not provided in the supplied evidence beyond the existence of guide RNA modifications.
The supplied evidence does not provide quantitative data on guide RNA performance, off-target rates, editing efficiency, or organism-specific constraints. It also notes that biomedical research and therapeutic applications of Cas9 were only beginning to be explored, indicating incomplete validation for translational use in the cited source.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Source 2primary paper2023Molecular Biology
Ranked Claims
Specificity of CRISPR/Cas9 can be modulated through guide RNA modifications.
possibilities to modulate their specificity through guide RNA modifications
Possible applications of Cas9 in biomedical research and therapeutics are only beginning to be explored.
With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics.
Possible applications of Cas9 in biomedical research and therapeutics are only beginning to be explored.
With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics.
Possible applications of Cas9 in biomedical research and therapeutics are only beginning to be explored.
With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics.
Possible applications of Cas9 in biomedical research and therapeutics are only beginning to be explored.
With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics.
Possible applications of Cas9 in biomedical research and therapeutics are only beginning to be explored.
With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics.
Possible applications of Cas9 in biomedical research and therapeutics are only beginning to be explored.
With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics.
Possible applications of Cas9 in biomedical research and therapeutics are only beginning to be explored.
With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics.
Cas9 is described as a powerful tool for engineering the genome in diverse organisms.
The Cas9 protein ... is emerging as a powerful tool for engineering the genome in diverse organisms.
Development of Cas9 as a tool made sequence-specific gene editing several magnitudes easier.
its development as a tool has made sequence-specific gene editing several magnitudes easier
Development of Cas9 as a tool made sequence-specific gene editing several magnitudes easier.
its development as a tool has made sequence-specific gene editing several magnitudes easier
Development of Cas9 as a tool made sequence-specific gene editing several magnitudes easier.
its development as a tool has made sequence-specific gene editing several magnitudes easier
Development of Cas9 as a tool made sequence-specific gene editing several magnitudes easier.
its development as a tool has made sequence-specific gene editing several magnitudes easier
Development of Cas9 as a tool made sequence-specific gene editing several magnitudes easier.
its development as a tool has made sequence-specific gene editing several magnitudes easier
Development of Cas9 as a tool made sequence-specific gene editing several magnitudes easier.
its development as a tool has made sequence-specific gene editing several magnitudes easier
Development of Cas9 as a tool made sequence-specific gene editing several magnitudes easier.
its development as a tool has made sequence-specific gene editing several magnitudes easier
Cas9 is an RNA-guided DNA endonuclease that can be reprogrammed to new target sites by changing the guide RNA sequence.
As an RNA-guided DNA endonuclease, Cas9 can be easily programmed to target new sites by altering its guide RNA sequence
Approval Evidence
2 sources2 linked approval claimsfirst-pass slug guide-rna
guide RNA modifications
Source:
Cas9 can be easily programmed to target new sites by altering its guide RNA sequence
Source:
specificity modulation strategysupports
Specificity of CRISPR/Cas9 can be modulated through guide RNA modifications.
possibilities to modulate their specificity through guide RNA modifications
Source:
mechanism or programmabilitysupports
Cas9 is an RNA-guided DNA endonuclease that can be reprogrammed to new target sites by changing the guide RNA sequence.
As an RNA-guided DNA endonuclease, Cas9 can be easily programmed to target new sites by altering its guide RNA sequence
Source:
Comparisons
Source-backed strengths
The main strength supported by the evidence is simple programmability: Cas9 can be easily targeted to new sites by altering guide RNA sequence. The literature further characterizes Cas9 as a powerful genome-engineering tool across diverse organisms, indicating broad utility of the guide RNA-directed targeting paradigm.
Source:
its development as a tool has made sequence-specific gene editing several magnitudes easier
Compared with antisense oligonucleotides
guide RNA and antisense oligonucleotides address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing
Strengths here: looks easier to implement in practice.
Compared with photo-sensitive circular gRNAs
guide RNA 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
Strengths here: looks easier to implement in practice.
Compared with synthetic riboswitches
guide RNA and synthetic riboswitches address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing
Relative tradeoffs: appears more independently replicated; looks easier to implement in practice.
Ranked Citations
- 1.