An intein-split transactivator for intersectional neural imaging and optogenetic manipulation.

Area of Science:

• Neuroscience
• Molecular Biology
• Genetics

Background:

• Cell-type-specific neural control is crucial for understanding brain function.
• Current methods for intersectional control have limitations in feature number, effector compatibility, and gene expression levels.

Purpose of the Study:

• To develop a novel technique for precise, multi-feature cell-type-specific gene expression control in mice.
• To overcome limitations of existing intersectional genetic tools.

Main Methods:

• Utilized protein-splicing mediated by intervening sequences (intein).
• Devised an intein-based intersectional synthesis of transactivator (IBIST) system.
• Validated IBIST for controlling fluorophores, optogenetic opsins, and Ca2+ indicators.

Main Results:

• IBIST demonstrated specificity and sufficiency in controlling gene expression across various intersectional conditions.
• IBIST successfully intersected up to five distinct cellular features.
• IBIST-based Ca2+ imaging revealed differential tuning of hippocampal neurons to emotional stimuli based on projection targets.

Conclusions:

• IBIST effectively multiplexes cell-type feature intersection and common effector control.
• IBIST offers a powerful tool for regulating gene expression, monitoring, and manipulating neural activity with high precision.