Recently I want to overexpress a gene in primarily cultured motor neuron. Someone told me if I use virus to infect the cells, I have to choose a motor neuron specific promoter because the gene will be inserted into the genome. But if i transiently transfect the gene using lipofectmine, it doesn't matter what promoter I choose. Is this true?
Hello,
For transfecting motor neurons, I recommend the NeuroMag transfection reagent that is based on the Magnetofection technology. It has been shown and published that 45% transfection efficiency can be achieved. For your convenience, you can find below references mentioning the use of this reagent to transfect motor neurons. The publication by C. Fallini et al. (Mol Neurodegener. 2010; 21(5):17) has described 45% transfection efficiency with NeuroMag in primary mouse motor neurons (see http://www.ncbi.nlm.nih.gov/pubmed/20406490). I recommend following the protocol defined in this paper. You can also find comments and the protocol on the Alzheimer Research Forum: http://www.alzforum.org/new/detail.asp?id=2461
In addition, other papers have been published on NeuroMag efficiency to transfect motor neurons.
http://www.ncbi.nlm.nih.gov/pubmed/25336041
http://www.ncbi.nlm.nih.gov/pubmed/22641816
http://www.ncbi.nlm.nih.gov/pubmed/21389246
http://www.ncbi.nlm.nih.gov/pubmed/20406490
http://www.ncbi.nlm.nih.gov/pubmed/21658607
http://onlinelibrary.wiley.com/doi/10.1002/ncn3.2/abstract
http://www.ncbi.nlm.nih.gov/pubmed/17475624
http://www.ncbi.nlm.nih.gov/pubmed/24920579
Good luck
Your idea is correct. To overexpress a protein, strong promoters like CMV are usually chosen. More specific promoters are used in situations like transgenic mice when DNA is inserted into the genome, but expression limited to a particular tissue is needed. If you are using a virus with cultured cells, strong promoters are still often used, but I suppose a specific promoter might be needed in some experiments. Reagents like lipofectamine usually produce poor transfection efficiency with primary cells, however, so you might need to use another method if you can't see expression. You could still use pCMV.
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