Source 1primary paper2023MED
Toolkit/single guide RNA array for multiplexed gene activation
single guide RNA array for multiplexed gene activation
Also known as: guide RNA array, single guide RNA array
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
This system allows versatile thermal induction of gene activation or base editing across cell types, including primary T cells, and enables multiplexed gene activation using a single guide RNA array.
Usefulness & Problems
Why this is useful
The guide RNA array enables multiplexed gene activation within the reported sonogenetic dCas12a platform. The abstract specifically states that multiplexed activation is achieved using a single guide RNA array.; multiplexed gene activation
Source:
The guide RNA array enables multiplexed gene activation within the reported sonogenetic dCas12a platform. The abstract specifically states that multiplexed activation is achieved using a single guide RNA array.
Source:
multiplexed gene activation
Problem solved
It addresses the need to activate multiple endogenous genes in a multiplexed manner. This is presented as a design feature of the reported platform.; supports regulation of multiple targets within one inducible CRISPR system
Source:
It addresses the need to activate multiple endogenous genes in a multiplexed manner. This is presented as a design feature of the reported platform.
Source:
supports regulation of multiple targets within one inducible CRISPR system
Problem links
supports regulation of multiple targets within one inducible CRISPR system
LiteratureIt addresses the need to activate multiple endogenous genes in a multiplexed manner. This is presented as a design feature of the reported platform.
Source:
It addresses the need to activate multiple endogenous genes in a multiplexed manner. This is presented as a design feature of the reported platform.
Published Workflows
Objective: Develop a precise, non-invasive, and tunable ultrasound-triggered platform for inducible endogenous gene regulation and base editing in vitro and in vivo.
Why it works: The abstract states that ultrasound absorption produces thermal energy, which activates a heat shock promoter controlling an engineered hyper-efficient dCas12a effector, thereby enabling inducible gene activation or base editing.
heat-shock-promoter-mediated inductionthermal activation from ultrasound absorptiondCas12a-based gene activationdCas12a-based base editingultrasound stimulationhigh-intensity focused ultrasoundguide RNA array multiplexing
Stages
- 1.Engineering of the ultrasound-responsive CRISPR effector system(library_design)
This stage creates the core inducible construct architecture that links ultrasound-derived heating to genome regulation or base editing.
Selection: Place a hyper-efficient dCas12a and effector under a heat shock promoter so the system can be activated by ultrasound-generated thermal energy.
- 2.Cross-cell-type functional testing(functional_characterization)
This stage establishes that the inducible system functions beyond a single cell context and includes primary T cells.
Selection: Test whether the system supports thermal induction of gene activation or base editing across cell types including primary T cells.
- 3.Multiplex activation demonstration with guide RNA array(secondary_characterization)
This stage shows that the platform can regulate multiple targets using the Cas12a-compatible guide-array design.
Selection: Demonstrate multiplexed gene activation using a single guide RNA array.
- 4.In vivo localized activation in mouse models(in_vivo_validation)
This stage tests whether the ultrasound-responsive system can be activated locally in vivo.
Selection: Use HIFU-guided localized temperature elevation to trigger reporter gene expression in implanted cells in mice.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Mechanisms
crispr-dcas12a-mediated multiplexed gene activationheat shock promoter-driven transcriptional activationthermal inductionultrasound-to-heat transductionTechniques
No technique tags yet.
Target processes
editingInput: Thermal
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: reporter
The abstract supports that this array is used together with the engineered hyper-efficient dCas12a effector system. It is part of the thermally inducible ultrasound-responsive setup.; used with the engineered dCas12a-based inducible system
The abstract does not state that the guide RNA array alone provides inducibility or delivery. It also does not provide target-count limits or comparative performance against other multiplexing formats.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
A single guide RNA array enables multiplexed gene activation in the reported system.
and enables multiplexed gene activation using a single guide RNA array
The paper presents an ultrasound-based strategy for precise, non-invasive, and tunable control of multiple endogenous genes in vitro and in vivo.
Here, we present a precise, non-invasive, and tunable strategy for controlling the expression of multiple endogenous genes both in vitro and in vivo, utilizing ultrasound as the stimulus.
The system supports thermal induction of gene activation or base editing across cell types including primary T cells.
This system allows versatile thermal induction of gene activation or base editing across cell types, including primary T cells
In mouse models, HIFU-guided localized temperature elevation triggers reporter gene expression in implanted cells.
In mouse models, localized temperature elevation guided by high-intensity focused ultrasound effectively triggers reporter gene expression in implanted cells.
The engineered system is inducibly activated by thermal energy produced through ultrasound absorption using a heat shock promoter and a hyper-efficient dCas12a effector.
By engineering a hyper-efficient dCas12a and effector under a heat shock promoter, we demonstrate a system that can be inducibly activated through thermal energy produced by ultrasound absorption.
Approval Evidence
1 source1 linked approval claimfirst-pass slug single-guide-rna-array-for-multiplexed-gene-activation
This system allows versatile thermal induction of gene activation or base editing across cell types, including primary T cells, and enables multiplexed gene activation using a single guide RNA array.
Source:
capabilitysupports
A single guide RNA array enables multiplexed gene activation in the reported system.
and enables multiplexed gene activation using a single guide RNA array
Source:
Comparisons
Source-backed strengths
enables multiplexed gene activation
Source:
enables multiplexed gene activation
Compared with CRISPR/Cas-hybrid assays
single guide RNA array for multiplexed gene activation and CRISPR/Cas-hybrid assays address a similar problem space because they share editing.
Shared frame: shared target processes: editing; same primary input modality: thermal
Strengths here: looks easier to implement in practice.
Compared with microfluidic organ-on-chip platforms
single guide RNA array for multiplexed gene activation and microfluidic organ-on-chip platforms 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 Z7-E78-ABE
single guide RNA array for multiplexed gene activation and Z7-E78-ABE address a similar problem space because they share editing.
Shared frame: same top-level item type; shared target processes: editing
Ranked Citations
- 1.