Highly efficient method for gene delivery into mouse dorsal root ganglia neurons
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Published on: March 18, 2013
Lingli Yu1, Florie Reynaud2, Julien Falk2
1Differentiation and Cell Cycle Group, Laboratoire de Biologie Moléculaire de la Cellule, UMR 5239, Centre National de la Recherche Scientifique, Ecole normale Supérieure de Lyon, University of Lyon 1 Claude Bernard, University of Lyon Lyon, France ; Laboratory of Molecular and Cellular Neurophysiology, East China Normal University Shanghai, China ; Joint Laboratory of Neuropathogenesis, Key Laboratory of Brain Functional Genomics, Chinese Ministry of Education, East China Normal University, Centre National de la Recherche Scientifique, Ecole Normale Supérieure de Lyon Shanghai, China.
Abstract:
The development of gene transfection technologies has greatly advanced our understanding of life sciences. While use of viral vectors has clear efficacy, it requires specific expertise and biological containment conditions. Electroporation has become an effective and commonly used method for introducing DNA into neurons and in intact brain tissue. The present study describes the use of the Neon® electroporation system to transfect genes into dorsal root ganglia neurons isolated from embryonic mouse Day 13.5-16. This cell type has been particularly recalcitrant and refractory to physical or chemical methods for introduction of DNA. By optimizing the culture condition and parameters including voltage and duration for this specific electroporation system, high efficiency (60-80%) and low toxicity (>60% survival) were achieved with robust differentiation in response to Nerve growth factor (NGF). Moreover, 3-50 times fewer cells are needed (6 × 10(4)) compared with other traditional electroporation methods. This approach underlines the efficacy of this type of electroporation, particularly when only limited amount of cells can be obtained, and is expected to greatly facilitate the study of gene function in dorsal root ganglia neuron cultures.
