Single-cell genetic manipulation is expected to substantially advance the field of systems neuroscience. This study examined the feasibility of combining in vivo whole-cell recording and gene delivery by answering four specific questions:
- Do cells survive for a substantial period after whole-cell recording in vivo?
- What is the likelihood and longevity of whole-cell recordings with internal solutions containing plasmid DNA?
- Can plasmid DNA dialyze into the cell and drive protein expression?
- Is protein expression restricted to the patched cell?
The delivery of DNA into neurons through an intracellular recording method should be considered useful for any experimental system including neuronal and non-neuronal cultures where cells can be maintained for extended periods of time. The method of DNA delivery by whole-cell recording described here provides a versatile tool for both assaying and manipulating single cells and probing the functional connectivity of specific local and long-range networks.
Click here to view the full text publication: “Transfection via whole-cell recording in vivo; bridging single-cell physiology, genetics and connectomics.” Ede A Rancz, Kevin M Franks, Martin K Schwarz, Bruno Pichler, Andreas T Schaefer, Troy W Margrie. Nature Neuroscience Feb 2011