RNA-programmable cell type monitoring and manipulation in the human cortex with CellREADR

Area of Science:

• Neuroscience
• Molecular Biology
• Genetics

Background:

• Understanding human brain circuits requires precise experimental access to diverse neuron types.
• Current methods for targeting human neural populations are limited, relying on transcriptional enhancers and viral capsids.

Purpose of the Study:

• To demonstrate CellREADR, a novel RNA sensor-effector technology, for accessing, monitoring, and manipulating specific human neuron types in ex vivo cortical tissues.
• To validate CellREADR's ability to target CALB2+ GABAergic interneurons and FOXP2+ glutamatergic projection neurons.

Main Methods:

• Designed CellREADR constructs to couple RNA sensing with effector protein translation for cell-type-specific targeting.
• Validated targeting using histological, electrophysiological, and transcriptomic analyses.
• Employed CellREADR to express optogenetic effectors and calcium indicators for neuronal manipulation and monitoring.

Main Results:

• Successfully demonstrated CellREADR's ability to specifically target and access distinct human neuron types (CALB2+ and FOXP2+).
• Validated cell targeting through comprehensive histological, electrophysiological, and transcriptomic methods.
• Showcased CellREADR-mediated manipulation and monitoring of targeted neuronal populations within human cortical microcircuits.
• Demonstrated the combined use of AAV-based CellREADR and enhancer vectors for co-targeting different subpopulations.

Conclusions:

• CellREADR provides specific, reliable, and programmable experimental access to targeted human cell types.
• This technology holds significant potential for advancing the study of human neural circuits.
• CellREADR offers a promising avenue for developing cell type-specific treatments for brain disorders.