Single-cell genetic manipulation is expected to substantially advance the field of systems neuroscience. However, existing gene delivery techniques do not allow researchers to electrophysiologically characterize cells and to thereby establish an experimental link between physiology and genetics for understanding neuronal function. In the mouse brain in vivo, we found that neurons remained intact after 'blind' whole-cell recording, that DNA vectors could be delivered through the patch-pipette during such recordings and that these vectors drove protein expression in recorded cells for at least 7 d. To illustrate the utility of this approach, we recorded visually evoked synaptic responses in primary visual cortical cells while delivering DNA plasmids that allowed retrograde, monosynaptic tracing of each neuron's presynaptic inputs. By providing a biophysical profile of a cell before its specific genetic perturbation, this combinatorial method captures the synaptic and anatomical receptive field of a neuron.
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K.M.F. thanks S. Siegelbaum and R. Axel for support and encouragement and A. Mulligan for technical assistance. We also thank P.H. Seeburg for support; D. Castro, B. Matynoga and D. Drechsel for advice; K.-K. Conzelmann for rabies virus SAD-ΔG-mCherry; and P. Boross, E. Callaway, H. Wildner and F. Guillemot for discussions and reagents. E.A.R. is a Sir Henry Wellcome Postdoctoral Fellow. This work was supported by The Robert Leet and Clara Guthrie Patterson Trust and a National Institute on Deafness and Other Communication Disorders (NIDCD) K99 grant (K.M.F.), The Max-Planck Gesellschaft (A.T.S., M.K.S.), the Medical Research Council (MRC) and The Alexander Von Humboldt Foundation (T.W.M.).
The authors declare no competing financial interests.
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Rancz, E., Franks, K., Schwarz, M. et al. Transfection via whole-cell recording in vivo: bridging single-cell physiology, genetics and connectomics. Nat Neurosci 14, 527–532 (2011). https://doi.org/10.1038/nn.2765
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