Fluorescently-labeled split-QF hemidrivers: simplifying and enhancing methods enabling intersectional targeting of...

Area of Science:

• Neuroscience
• Genetics
• Molecular Biology

Background:

• Single-cell transcriptomics reveals complex cell type diversity defined by gene expression codes.
• Intersectional targeting strategies are crucial for selective cell labeling and manipulation.
• Split-driver systems (e.g., Gal4/UAS, QF/QUAS) are effective in invertebrates but underutilized in vertebrates.

Purpose of the Study:

• To engineer fluorescently labeled split-QF and QF2 hemidrivers for vertebrate systems.
• To enhance the specificity and ease of use of intersectional targeting tools.
• To facilitate functional studies of cell types identified by single-cell transcriptomics.

Main Methods:

• Development of fluorescently labeled split-QF and QF2 hemidrivers.
• Utilized direct reporter fusions and the 2A viral peptide co-expression system.
• Incorporated fluorescent labeling for simplified line creation, direct visualization, and quality control.

Main Results:

• Successfully engineered fluorescently labeled split-QF and QF2 hemidrivers in zebrafish.
• Fluorescent labeling simplifies hemidriver transgenic line creation and maintenance.
• Enabled direct visualization of Activation Domain (AD) and DNA Binding Domain (DBD) intersects and robust quality control.

Conclusions:

• Engineered fluorescently labeled split-QF and -QF2 hemidrivers in zebrafish.
• These tools simplify the generation of intersectional targeting tools.
• Facilitate robust quality control assessments of split-driver function for enhanced transgenic targeting specificity.