Nucleofection of primary neurons
Efficient gene transfer is an important tool for the study of neuronal function and biology. This has proved difficult and inefficient with traditional transfection strategies, which can also be fairly toxic, whereas viral-mediated gene transfer, although highly efficient, is often time-consuming. The recently developed Amaxa Nucleofector technology, based on electroporation with preset parameters in a cell-type-specific solution, enables direct delivery of DNA, small interfering (si)RNA oligonucleotides and siRNA vectors into the cell nucleus. This strategy results in reproducible, rapid, and efficient transfection of a broad range of cells, including primary neurons. Nucleofected neurons survive for up to 3 weeks and remain functional. We are currently using this transfection method to examine the contribution of Rho GTPase signaling pathways in the establishment of neuronal polarity, neuronal migration, and neurite outgrowth. Here, we describe three protocols to efficiently nucleofect rat cerebellar granule, cortical, and hippocampal neurons.
| Medienart: |
Artikel |
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| Erscheinungsjahr: |
2006 |
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| Erschienen: |
2006 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:406 |
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| Enthalten in: |
Methods in enzymology - 406(2006) vom: 30., Seite 374-88 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Gärtner, Annette [VerfasserIn] |
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| Themen: |
Journal Article |
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| Anmerkungen: |
Date Completed 23.03.2006 Date Revised 13.08.2007 published: Print Citation Status MEDLINE |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM160665310 |
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| 245 | 1 | 0 | |a Nucleofection of primary neurons |
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| 500 | |a Date Revised 13.08.2007 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Efficient gene transfer is an important tool for the study of neuronal function and biology. This has proved difficult and inefficient with traditional transfection strategies, which can also be fairly toxic, whereas viral-mediated gene transfer, although highly efficient, is often time-consuming. The recently developed Amaxa Nucleofector technology, based on electroporation with preset parameters in a cell-type-specific solution, enables direct delivery of DNA, small interfering (si)RNA oligonucleotides and siRNA vectors into the cell nucleus. This strategy results in reproducible, rapid, and efficient transfection of a broad range of cells, including primary neurons. Nucleofected neurons survive for up to 3 weeks and remain functional. We are currently using this transfection method to examine the contribution of Rho GTPase signaling pathways in the establishment of neuronal polarity, neuronal migration, and neurite outgrowth. Here, we describe three protocols to efficiently nucleofect rat cerebellar granule, cortical, and hippocampal neurons | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 700 | 1 | |a Collin, Ludovic |e verfasserin |4 aut | |
| 700 | 1 | |a Lalli, Giovanna |e verfasserin |4 aut | |
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