HiFENS

HiFENS enables high-throughput detection of endogenous alternative pre-mRNA splicing isoforms using hybridization chain reaction (HCR) to support single-cell-resolution functional genomics analyses.

Key Features:

• High-Throughput Capability: Enables simultaneous examination of multiple genes and their splicing isoforms.
• Specificity and Sensitivity: Provides optimized detection with high specificity for accurate identification of endogenous splicing isoforms.
• Multiplexing Ability: Supports multiplexing of probes to detect various splicing outcomes within individual cells.
• Single-Cell Resolution: Detects splicing events at single-cell resolution to analyze cellular heterogeneity in isoform expression.
• HCR Amplification: Uses hybridization chain reaction (HCR) to amplify RNA signals from specific targets in situ.
• Functional Genomics Screening: Designed to support functional genomics screens such as RNAi-based screens of candidate regulators.

Scientific Applications:

• Functional Genomics Screening: Enables high-throughput screens to identify regulators of alternative splicing.
• Regulatory Mechanism Mapping: Facilitates investigation of upstream factors and signal transduction pathways that influence splicing factor activity.
• Single-Cell Isoform Analysis: Characterizes cell-to-cell variability in splicing isoform expression relevant to biological processes and disease.

Methodology:

Uses hybridization chain reaction (HCR) to amplify and visualize RNA splicing isoforms in situ and was applied in an RNAi screen targeting 702 human kinases to identify regulators of FGFR2 alternative splicing.