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In utero ventricular microinjection and electroporation
[Describe principle, how it compares to other techniques in terms of expression levels, what brain areas are amenable to in utero electroporation, protocol]
This technique was used successfully to express ChR2 in the following papers:
- Huber D, Petreanu L, Ghitani N, Ranade S, Hromádka T, Mainen Z, Svoboda K. Sparse optical microstimulation in barrel cortex drives learned behaviour in freely moving mice. Nature. 2008 Jan 3;451(7174):61-4. In this paper, ChR2–GFP was electroporated into layer 2/3 progenitor cells in mice embryos at E16.
- Sparse transgene expression in the cortex and hippocampus using the Thy1S promoter and in utero electroporation .
- In Utero Intraventricular Injection and Electroporation of E15 Mouse Embryos from the Journal of Visualized Experiments.
Postnatal non-ventricular microinjection and electroporation
- Gene Delivery to Postnatal Rat Brain by Non-ventricular Plasmid Injection and Electroporation from the Journal of Visualized Experiments.
The Häusser lab has developed a protocol for electroporating single of groups of neurons in vivo. This methods allows labeling very few neurons at precise locations within the brain. Satisfactory expression levels (in particular for ChR2 expression) appear to be reached only after 48 hours.
- Judkewitz B, Rizzi M, Kitamura K, Häusser M. Targeted single-cell electroporation of mammalian neurons in vivo. Nat Protoc. 2009;4(6):862-9.
- Kitamura K, Judkewitz B, Kano M, Denk W, Häusser M. Targeted patch-clamp recordings and single-cell electroporation of unlabeled neurons in vivo. Nat Methods. 2008 Jan;5(1):61-7.
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