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A novel method for RNA interference in neurons using enhanced green fluorescent protein (EGFP)-transgenic rats |
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| Authors: | Lu Jia Nozumi Motohiro Fujii Hiroshi Igarashi Michihiro |
| Affiliation: | aDivision of Molecular and Cellular Biology, Graduate School of Medical and Dental Sciences, Niigata University, Asahi-machi, Chuo-ku, Niigata 951-8510, Japan bDivision of Ophthalmology, Graduate School of Medical and Dental Sciences, Niigata University, Asahi-machi, Chuo-ku, Niigata 951-8510, Japan cTrans-disciplinary Research Programs, Niigata University, Asahi-machi, Chuo-ku, Niigata 951-8510, Japan dDepartment of Ophthalmology, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province 150001, PR China eDepartment of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, Minamiminowa, Nagano 399-4598, Japan |
| Abstract: | RNA interference (RNAi) is the simplest way of examining gene function by inhibiting expression. However, due to the low rate of introducing short interfering RNA (siRNA) into neurons, it is difficult to discriminate into which neurons that have been successfully introduced. Here, we used neurons from transgenic rats expressing enhanced green fluorescent protein (EGFP), and we simultaneously applied small interfering RNAs (siRNAs) against EGFP and a targeted gene to the EGFP-expressing neurons. EGFP fluorescence and immunoreactivity of the protein were then assessed by immunofluorescence microscopy. Quantitative analysis of the immunofluorescence confirmed that loss of EGFP closely correlates with loss of the target protein. These results indicate that this method can be used in a wider range of the neuroscientific research, especially in genome-wide studies. |
| Keywords: | Short interfering RNA (siRNA) Immunofluorescence Enhanced green fluorescent protein (EGFP) Growth cone Axonal growth Transgenic mice Fatty acid-binding protein (FABP) |
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