Source 1primary paper2025MED
Toolkit/lentiviral delivery of pegRNAs
lentiviral delivery of pegRNAs
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
lentiviral delivery of pegRNAs, ensuring robust, ubiquitous, and sustained expression of both prime editors and pegRNAs
Usefulness & Problems
No literature-backed usefulness or problem-fit explainer has been materialized for this record yet.
Published Workflows
Objective: Maximize prime editing efficiency and versatility across diverse cell types by systematically optimizing editor expression, pegRNA delivery, and related implementation variables.
Why it works: The abstract states that the combined strategy ensures robust, ubiquitous, and sustained expression of both prime editors and pegRNAs, which is presented as the basis for improved editing efficiency.
stable genomic integration of prime editorssustained expression of prime editors and pegRNAssystematic optimizationpiggyBac-mediated integrationsingle-clone selectionpromoter optimizationlentiviral pegRNA delivery
Stages
- 1.system optimization and build(library_build)
This stage exists to implement the optimization stack intended to maximize prime editing efficiency.
Selection: Combine stable editor integration, single-clone selection, enhanced promoter use, and lentiviral pegRNA delivery to improve prime editing performance.
- 2.performance assessment across loci and cell lines(functional_characterization)
This stage exists to determine whether the systematic optimizations improve editing efficiency broadly across test settings.
Selection: Assess editing efficiency across multiple loci and cell lines.
- 3.validation in challenging hPSC contexts(confirmatory_validation)
This stage exists to assess whether the optimized system remains effective in challenging cell types beyond the initial cell-line settings.
Selection: Test the optimized system in primed and naïve human pluripotent stem cells.
Steps
- 1.stably integrate prime editors using piggyBaceditor integration harness
Establish stable genomic integration of prime editors.
Stable editor integration is part of the optimization stack used to support robust and sustained expression before downstream validation.
- 2.select integrated single clones
Isolate single clones carrying the integrated editor system.
This follows editor integration because clone selection depends on having integrated cells to choose from.
- 3.use an enhanced promoter
Increase expression performance within the optimized prime editing system.
Promoter choice is part of the expression-optimization stack used to support robust and ubiquitous expression before performance testing.
- 4.deliver pegRNAs by lentiviruspegRNA delivery harness
Provide pegRNAs with robust, ubiquitous, and sustained expression.
PegRNA delivery is included as a key delivery optimization before evaluating editing outcomes.
- 5.measure editing efficiency across multiple loci and cell lines
Assess whether the optimization stack improves prime editing performance broadly.
Performance testing follows system construction so the combined optimizations can be evaluated.
- 6.validate the optimized system in primed and naïve hPSCs
Test whether the optimized system remains effective in challenging cell types.
This confirmatory validation is performed after broader cell-line testing to further assess efficacy in a more challenging context.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A delivery strategy grouped with the mechanism branch because it determines how a system is instantiated and deployed in context.
Techniques
Selection / EnrichmentTarget processes
selectionValidation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
The optimized approach combined piggyBac-mediated stable prime editor integration, single-clone selection, an enhanced promoter, and lentiviral pegRNA delivery to ensure robust, ubiquitous, and sustained expression of prime editors and pegRNAs.
our approach combined stable genomic integration of prime editors via the piggyBac transposon system, selection of integrated single clones, the use of an enhanced promoter, and lentiviral delivery of pegRNAs, ensuring robust, ubiquitous, and sustained expression of both prime editors and pegRNAs
A systematically optimized prime editing strategy achieved up to 80% editing efficiency across multiple loci and cell lines.
we implemented a series of systematic optimizations, achieving up to 80% editing efficiency across multiple loci and cell lines
editing efficiency 80 %
The optimized prime editing strategy provides a highly efficient and versatile framework for in vitro genome engineering.
our optimized prime editing strategy provides a highly efficient and versatile framework for genome engineering in vitro
The optimized prime editing system achieved up to 50% editing efficiency in human pluripotent stem cells in both primed and naïve states.
we validated our optimized system in human pluripotent stem cells (hPSCs) in both primed and naïve states, achieving substantial editing efficiencies of up to 50%
editing efficiency 50 %
Approval Evidence
1 source1 linked approval claimfirst-pass slug lentiviral-delivery-of-pegrnas
lentiviral delivery of pegRNAs, ensuring robust, ubiquitous, and sustained expression of both prime editors and pegRNAs
Source:
mechanistic rationalesupports
The optimized approach combined piggyBac-mediated stable prime editor integration, single-clone selection, an enhanced promoter, and lentiviral pegRNA delivery to ensure robust, ubiquitous, and sustained expression of prime editors and pegRNAs.
our approach combined stable genomic integration of prime editors via the piggyBac transposon system, selection of integrated single clones, the use of an enhanced promoter, and lentiviral delivery of pegRNAs, ensuring robust, ubiquitous, and sustained expression of both prime editors and pegRNAs
Source:
Comparisons
No literature-backed comparison notes have been materialized for this record yet.
Ranked Citations