Source 1primary paper2024MED
Toolkit/drug-selectable acoustic reporter gene system
drug-selectable acoustic reporter gene system
Also known as: inducible and drug-selectable acoustic reporter gene system
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Here, we describe an inducible and drug-selectable acoustic reporter gene system that can enable gas vesicle expression in mammalian cell lines, which we demonstrate using HEK293T cells.
Usefulness & Problems
Why this is useful
This system is an inducible, drug-selectable acoustic reporter gene construct design for generating gas-vesicle-expressing mammalian cells that can be imaged by ultrasound. The abstract presents it as a reporter-gene platform for visualizing specific cell types.; enabling gas vesicle expression in mammalian cell lines; ultrasound imaging of specific cell types; reducing cell processing time when establishing gas-vesicle-expressing mammalian cells
Source:
This system is an inducible, drug-selectable acoustic reporter gene construct design for generating gas-vesicle-expressing mammalian cells that can be imaged by ultrasound. The abstract presents it as a reporter-gene platform for visualizing specific cell types.
Source:
enabling gas vesicle expression in mammalian cell lines
Source:
ultrasound imaging of specific cell types
Source:
reducing cell processing time when establishing gas-vesicle-expressing mammalian cells
Problem solved
It addresses the long processing time and instability associated with establishing mammalian cell lines that robustly express gas vesicles for ultrasound contrast. The design is said to increase the fraction of cells integrating all plasmids and to make mixed selected populations usable.; low stability and low proportion of mammalian cells successfully integrating all required plasmids; need for lengthy processing to establish clonal or polyclonal gas-vesicle-expressing mammalian cell lines
Source:
It addresses the long processing time and instability associated with establishing mammalian cell lines that robustly express gas vesicles for ultrasound contrast. The design is said to increase the fraction of cells integrating all plasmids and to make mixed selected populations usable.
Source:
low stability and low proportion of mammalian cells successfully integrating all required plasmids
Source:
need for lengthy processing to establish clonal or polyclonal gas-vesicle-expressing mammalian cell lines
Problem links
low stability and low proportion of mammalian cells successfully integrating all required plasmids
LiteratureIt addresses the long processing time and instability associated with establishing mammalian cell lines that robustly express gas vesicles for ultrasound contrast. The design is said to increase the fraction of cells integrating all plasmids and to make mixed selected populations usable.
Source:
It addresses the long processing time and instability associated with establishing mammalian cell lines that robustly express gas vesicles for ultrasound contrast. The design is said to increase the fraction of cells integrating all plasmids and to make mixed selected populations usable.
need for lengthy processing to establish clonal or polyclonal gas-vesicle-expressing mammalian cell lines
LiteratureIt addresses the long processing time and instability associated with establishing mammalian cell lines that robustly express gas vesicles for ultrasound contrast. The design is said to increase the fraction of cells integrating all plasmids and to make mixed selected populations usable.
Source:
It addresses the long processing time and instability associated with establishing mammalian cell lines that robustly express gas vesicles for ultrasound contrast. The design is said to increase the fraction of cells integrating all plasmids and to make mixed selected populations usable.
Published Workflows
Objective: Establish a mammalian acoustic reporter gene system that enables gas vesicle expression with less cell processing and sufficient ultrasound contrast from a drug-selected mixed population.
Why it works: The abstract states that the drug-selectable construct design increases the stability and proportion of cells that successfully integrate all plasmids into their genome, which is presented as the reason cell processing time can be reduced while retaining robust ultrasound contrast.
gas vesicle expression in mammalian cellsgenetically encoded ultrasound contrastinducible expressiondrug selection
Stages
- 1.Drug-selectable construct integration and enrichment(selection)
This stage exists to increase the stability and proportion of cells with all required plasmids integrated, reducing the need for more labor-intensive downstream isolation steps.
Selection: Drug selection enriches for cells that successfully integrate all plasmids into their genome.
- 2.Bypass of single-cell cloning and fluorescence-activated cell sorting(decision_gate)
This decision point narrows the workflow by avoiding additional cell-isolation steps when the selected mixed population already meets the reported imaging objective.
- 3.Optical and ultrasound verification(confirmatory_validation)
This stage confirms that the selected engineered cells actually express gas vesicles and produce measurable acoustic contrast.
Selection: Verification of successful gas vesicle expression and ultrasound contrast relative to controls.
Steps
- 1.Use inducible drug-selectable construct design to enrich for cells integrating all plasmidsengineered construct system
Increase the stability and proportion of cells that successfully integrate all plasmids into their genome.
This enrichment step is presented as the mechanism for reducing later cell processing burden.
- 2.Proceed with drug-selected mixed population instead of single-cell cloning or FACSselected engineered cell population
Avoid additional cell processing steps while retaining sufficient ultrasound contrast.
The abstract states this decision is justified after drug selection because the mixed population is sufficient to generate robust ultrasound contrast.
- 3.Verify gas vesicle expression optically and ultrasonicallyengineered reporter system under test
Confirm successful gas vesicle expression and quantify ultrasound contrast relative to controls.
This confirmatory assay follows selection and workflow simplification to establish that the resulting population still produces the intended imaging phenotype.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Mechanisms
acoustic contrast generation by gas vesicle expressionchemical inductiondrug selectiongenomic integration of multiple plasmidsTechniques
Computational DesignTarget processes
recombinationInput: Chemical
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: reporter
The abstract indicates that implementation requires mammalian cell lines, plasmid integration into the genome, inducible control, drug selection, and optical and ultrasound verification. It is demonstrated in HEK293T cells.; requires mammalian cell engineering with genome integration of all plasmids; requires drug selection; requires inducible expression
The abstract does not show performance beyond the demonstrated mammalian cell context or specify exact construct components. It also does not establish in vivo or therapeutic efficacy in the abstract.; demonstrated in HEK293T cells in the abstract
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
This technology can be used to visualize specific cell types with ultrasound for applications including cellular reporting and cell therapies.
This technology presents a new reporter gene paradigm by which ultrasound can be harnessed to visualize specific cell types for applications including cellular reporting and cell therapies.
An inducible and drug-selectable acoustic reporter gene system can enable gas vesicle expression in mammalian cell lines.
Here, we describe an inducible and drug-selectable acoustic reporter gene system that can enable gas vesicle expression in mammalian cell lines, which we demonstrate using HEK293T cells.
The drug-selectable construct design increases the stability and proportion of cells that successfully integrate all plasmids into their genome, reducing the amount of cell processing time required.
Our drug-selectable construct design increases the stability and proportion of cells that successfully integrate all plasmids into their genome, thus reducing the amount of cell processing time required.
Cells expressing gas vesicles exhibited greater ultrasound signal-to-noise ratio than controls.
Successful gas vesicle expression was optically and ultrasonically verified, with cells expressing gas vesicles exhibiting an 80% greater signal-to-noise ratio compared to negative controls and a 500% greater signal-to-noise ratio compared to wild-type HEK293T cells.
signal-to-noise ratio increase versus negative controls 80 %signal-to-noise ratio increase versus wild-type HEK293T cells 500 %
The drug-selectable strategy eliminates the need for single-cell cloning and fluorescence-activated cell sorting, and a drug-selected mixed population is sufficient to generate robust ultrasound contrast.
Additionally, we demonstrate that our drug-selectable strategy forgoes the need for single-cell cloning and fluorescence-activated cell sorting, and that a drug-selected mixed population is sufficient to generate robust ultrasound contrast.
Approval Evidence
1 source5 linked approval claimsfirst-pass slug drug-selectable-acoustic-reporter-gene-system
Here, we describe an inducible and drug-selectable acoustic reporter gene system that can enable gas vesicle expression in mammalian cell lines, which we demonstrate using HEK293T cells.
Source:
application scopesupports
This technology can be used to visualize specific cell types with ultrasound for applications including cellular reporting and cell therapies.
This technology presents a new reporter gene paradigm by which ultrasound can be harnessed to visualize specific cell types for applications including cellular reporting and cell therapies.
Source:
capabilitysupports
An inducible and drug-selectable acoustic reporter gene system can enable gas vesicle expression in mammalian cell lines.
Here, we describe an inducible and drug-selectable acoustic reporter gene system that can enable gas vesicle expression in mammalian cell lines, which we demonstrate using HEK293T cells.
Source:
engineering advantagesupports
The drug-selectable construct design increases the stability and proportion of cells that successfully integrate all plasmids into their genome, reducing the amount of cell processing time required.
Our drug-selectable construct design increases the stability and proportion of cells that successfully integrate all plasmids into their genome, thus reducing the amount of cell processing time required.
Source:
performancesupports
Cells expressing gas vesicles exhibited greater ultrasound signal-to-noise ratio than controls.
Successful gas vesicle expression was optically and ultrasonically verified, with cells expressing gas vesicles exhibiting an 80% greater signal-to-noise ratio compared to negative controls and a 500% greater signal-to-noise ratio compared to wild-type HEK293T cells.
Source:
workflow simplificationsupports
The drug-selectable strategy eliminates the need for single-cell cloning and fluorescence-activated cell sorting, and a drug-selected mixed population is sufficient to generate robust ultrasound contrast.
Additionally, we demonstrate that our drug-selectable strategy forgoes the need for single-cell cloning and fluorescence-activated cell sorting, and that a drug-selected mixed population is sufficient to generate robust ultrasound contrast.
Source:
Comparisons
Source-stated alternatives
The abstract contrasts this strategy with workflows requiring single-cell cloning and fluorescence-activated cell sorting. It also contrasts it with prior methods for producing gas-vesicle-expressing mammalian cell lines that require substantial cell processing time.
Source:
The abstract contrasts this strategy with workflows requiring single-cell cloning and fluorescence-activated cell sorting. It also contrasts it with prior methods for producing gas-vesicle-expressing mammalian cell lines that require substantial cell processing time.
Source-backed strengths
increases stability and proportion of cells that successfully integrate all plasmids; reduces cell processing time required; forgoes the need for single-cell cloning; forgoes the need for fluorescence-activated cell sorting; drug-selected mixed populations are reported as sufficient for robust ultrasound contrast
Source:
increases stability and proportion of cells that successfully integrate all plasmids
Source:
reduces cell processing time required
Source:
forgoes the need for single-cell cloning
Source:
forgoes the need for fluorescence-activated cell sorting
Source:
drug-selected mixed populations are reported as sufficient for robust ultrasound contrast
Compared with fluorescence-activated cell sorting
The abstract contrasts this strategy with workflows requiring single-cell cloning and fluorescence-activated cell sorting. It also contrasts it with prior methods for producing gas-vesicle-expressing mammalian cell lines that require substantial cell processing time.
Shared frame: source-stated alternative in extracted literature
Strengths here: increases stability and proportion of cells that successfully integrate all plasmids; reduces cell processing time required; forgoes the need for single-cell cloning.
Relative tradeoffs: demonstrated in HEK293T cells in the abstract.
Source:
The abstract contrasts this strategy with workflows requiring single-cell cloning and fluorescence-activated cell sorting. It also contrasts it with prior methods for producing gas-vesicle-expressing mammalian cell lines that require substantial cell processing time.
Compared with single-cell transcriptomics
The abstract contrasts this strategy with workflows requiring single-cell cloning and fluorescence-activated cell sorting. It also contrasts it with prior methods for producing gas-vesicle-expressing mammalian cell lines that require substantial cell processing time.
Shared frame: source-stated alternative in extracted literature
Strengths here: increases stability and proportion of cells that successfully integrate all plasmids; reduces cell processing time required; forgoes the need for single-cell cloning.
Relative tradeoffs: demonstrated in HEK293T cells in the abstract.
Source:
The abstract contrasts this strategy with workflows requiring single-cell cloning and fluorescence-activated cell sorting. It also contrasts it with prior methods for producing gas-vesicle-expressing mammalian cell lines that require substantial cell processing time.
Ranked Citations
- 1.