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Georg Kosche @gek0s.bsky.social · 15/09/2025

We are excited by the potential benefits of single-cell RNA electroporation in elucidating the functions of hard-to-study ion channels or cell-type specific proteins in vivo, with the ability to precisely pre-select the neurons to be edited or modified.

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Georg Kosche @gek0s.bsky.social · 15/09/2025

Using whole-cell recordings in awake animals, we discovered a sequential program of adaptation to the disruption of E/I balance: After an initial increase in spiking and reduction in excitatory input (2-3d), neurons ultimately silenced themselves (7d+) through changes in membrane excitability.

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Georg Kosche @gek0s.bsky.social · 15/09/2025

We used this method to investigate how single L2/3 neurons respond to a loss of fast synaptic inhibition in vivo, when the rest of the network is unaffected. We delivered CRISPR/Cas9 against an essential subunit of the GABA-A receptor, which led to a reliable loss of inhibition in targeted cells.

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Georg Kosche @gek0s.bsky.social · 15/09/2025

We found RNA based single-cell electroporation to allow for high success rates (>85%) in protein expression as early as day 1 after electroporation. Success rate does not decrease when using many RNAs for CRISPR/Cas9 editing, resulting in very reliable in vivo gene editing.

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Georg Kosche @gek0s.bsky.social · 15/09/2025

Single-cell electroporation is performed via patch pipette and under 2-photon visual guidance in vivo. A short voltage pulse train is applied through the pipette to deliver RNA molecules into a targeted cell. The video below shows this process in cortical neurons expressing GCaMP6s.

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