Transfection of Neuronal Cultures.

Szczegóły
Abstrakt

Tytuł:: Transfection of Neuronal Cultures.
Autorzy:: Sariyer IK; Department of Neuroscience, Lewis Katz School of Medicine, Temple University, Philadelphia, PA, USA. .
Źródło:: Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2021; Vol. 2311, pp. 147-153.
Typ publikacji:: Journal Article; Research Support, N.I.H., Extramural
Język:: English
Imprint Name(s):: Publication: Totowa, NJ : Humana Press
Original Publication: Clifton, N.J. : Humana Press,
MeSH Terms:: Transfection*
Lipids/*chemistry
Neurons/*physiology
Animals ; Cells, Cultured ; Gene Expression Regulation ; Humans ; Neurons/metabolism ; Primary Cell Culture
References:: http://www.promega.com/resources/product-guides-and-selectors/protocols-and-applications-guide/transfection/.
Zeitelhofer M, John P, Vessey JP, Thomas S, Kiebler M, Dahm R (2009) Transfection of cultured primary neurons via nucleofection. Curr Protoc Neurosci Chapter 4:Unit4.32. (PMID: 19340811)
Karra D, Dahm R (2010) Transfection Techniques for Neuronal Cells. J Neurosci 30(18):6171–6177. (PMID: 10.1523/JNEUROSCI.0183-10.2010)
Washbourne P, McAllister AK (2002) Techniques for gene transfer into neurons. Curr Opin Neurobiol 12:566–573. (PMID: 10.1016/S0959-4388(02)00365-3)
Dib-Hajj SD, Choi JS, Macala LJ, Tyrrell L, Black JA, Cummins TR, Waxman SG (2009) Transfection of rat or mouse neurons by biolistics or electroporation. Nat Protoc 4:1118–1126. (PMID: 10.1038/nprot.2009.90)
Gresch O, Engel FB, Nesic D, Tran TT, England HM, Hickman ES, Korner I, Gan L, Chen S, Castro-Obregon S, Hammermann R, Wolf J, Müller-Hartmann H, Nix M, Siebenkotten G, Kraus G, Lun K (2004) New non-viral method for gene transfer into primary cells. Methods 33:151–163. (PMID: 10.1016/j.ymeth.2003.11.009)
Gartner A, Collin L, Lalli G (2006) Nucleofection of primary neurons. Methods Enzymol 406:374–388. (PMID: 10.1016/S0076-6879(06)06027-7)
Chen C, Okayama H (1987) High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol 7:2745–2752. (PMID: 10.1128/MCB.7.8.2745)
Yu LY, Arumäe U (2008) Survival assay of transiently transfected dopaminergic neurons. J Neurosci Methods 169(1):8–15. (PMID: 10.1016/j.jneumeth.2007.11.018)
Gabellini N, Minozzi MC, Leon A, Dal Toso R (1992) Nerve growth factor transcriptional control of c-fos promoter transfected in cultured spinal sensory neurons. J Cell Biol 118(1):131–138. (PMID: 10.1083/jcb.118.1.131)
Goetze B, Grunewald B, Baldassa S, Kiebler M (2004) Chemically controlled formation of a DNA/calcium phosphate coprecipitate: application for transfection of mature hippocampal neurons. J Neurobiol 60:517–525. (PMID: 10.1002/neu.20073)
Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Northrop JP, Ringold GM, Danielsen M (1987) Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci 84:7413–7417. (PMID: 10.1073/pnas.84.21.7413)
Felgner JH, Kumar R, Sridhar CN, Wheeler CJ, Tsai YJ, Border R, Ramsey P, Martin M, Felgner PL (1994) Enhanced gene delivery and mechanism studies with a novel series of cationic lipid formulations. J Biol Chem 269(4):2550–2561. (PMID: 10.1016/S0021-9258(17)41980-6)
Wiesenhofer B, Humpel C (2000) Lipid-mediated gene transfer into primary neurons using FuGene: comparison to C6 Glioma cells and primary glia. Exp Neurol 164:38–44. (PMID: 10.1006/exnr.2000.7414)
Ohki EC, Tilkins ML, Ciccarone VC, Price PJ (2001) Improving the transfection efficiency of post-mitotic neurons. J Neurosci Methods 112:95–99. (PMID: 10.1016/S0165-0270(01)00441-1)
Contributed Indexing:: Keywords: Calcium coprecipitation; Electroporation; FuGENE-6; Gene delivery; Lipofection; Nucleofection; Primary neurons; Transfection
Substance Nomenclature:: 0 (FuGene)
0 (Lipids)
Entry Date(s):: Date Created: 20210525 Date Completed: 20210811 Latest Revision: 20210811
Update Code:: 20240105
DOI:: 10.1007/978-1-0716-1437-2_10
PMID:: 34033082
: Czasopismo naukowe

Efficient transfection of genes into the neurons is a crucial step for the study of neuronal cell biology and functions. These include but not limited to investigating gene function by overexpression of target proteins via expression plasmids and knocking down the expression levels of neuronal genes by RNA interference (RNAi). In addition, reporter gene constructs are widely used to investigate the promoter activities of neuronal genes. Numerous transfection techniques have been established to deliver genes into the cells. However, efficient transfection of postmitotic cells, including neurons, still remains a challenging task. Here, we overview the advantages and disadvantages of various techniques for the transfection of primary neurons, and provide an optimized protocol for FuGENE-6 (Promega) which allows for a suitable transfection efficiency of primary neuronal cultures.

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