Summary
This project developed and validated scCHyMErA-Seq, an advanced single-cell platform for efficient combinatorial genetic perturbations and precise gene segment deletions, enabling simultaneous capture of guide RNAs and transcripts.
Establishing Combinatorial Screening Platform Coupled to Single-cell Analysis
US · IL
NIH
grant
awarded
#nih-1ZIABC012101-04
What they want
The project involved optimizing the hgRNA scaffold sequence and refining library preparation protocols to enhance the CHyMErA exon deletion screening platform for single-cell experiments. This advanced tool supports the simultaneous capture of Cas9 and Cas12a guide RNAs alongside polyadenylated transcripts at single-cell resolution. Validation included performing a screen focused on the deletion of alternative exons and applying scCHyMErA-Seq to high-throughput profiling of alternative cassette exons to identify regulatory effects on gene expression.
Technical requirements
- Optimization of hgRNA scaffold sequence
- Refinement of library preparation protocols
- Simultaneous capture of Cas9 and Cas12a guide RNAs
- Simultaneous capture of polyadenylated transcripts at single-cell resolution
- Efficient combinatorial genetic perturbations
- Precise gene segment deletions
- High-throughput profiling of alternative cassette exons
