Transfection generally refers to the introduction of foreign DNA into bacterial and/or mammalian cells. Transfection is an important tool used in studies investigating gene function and the modulation of gene expression, thus contributing to the advancement of basic cellular research, drug discovery, and target validation. This section provides an overview of different transfection methods, transfection workflow, factors affecting transfection efficient, and protocols.
Related Topics: Posttransfection Analysis of Cells, Instrument-Based Transfection Methods, and Chemical and Viral Transfection.
Transfection can be accomplished using chemical, biological, or physical methods. Common methods include electroporation, the use of a virus vector, lipofection, and biolistics. Many types of genetic material, including plasmid DNA, siRNA, proteins, dyes, and antibodies, may be transfected using any of these methods. However, a single method cannot be applied to all types of cells; transfection efficiencies and cytotoxicity may vary dramatically and depend on the method, cell type being utilized, and types of experiments being performed. Therefore, to obtain high efficiencies, all relevant factors should be considered for planning and selecting the appropriate transfection method.
Successful transfection is usually measured in terms of transfection efficiency and cell viability — the higher the efficiency and viability, the better the transfection. Several important factors, such as the DNA quantity and quality, cell type, cell health, and transfection method (as stated above) affect transfection results. Transfection efficiency, for example, varies greatly with the cell type and its physiological condition prior to transfection. Ideally, the cells should be actively growing, healthy, and free of contamination.
Another way to present factors impacting transfection results is to consider the entire transfection workflow and the series of key sub-experiments it comprises. Each sub-experiment can significantly affect efficiency and viability. The typical workflow for a transfection experiment is as follows:
Typical workflow for a transfection experiment.
The following table summarizes the factors to consider for efficient transfection:
Cells should be grown in medium appropriate for the cell line, supplemented with serum or growth factors as needed for viability
Transfect cells at 40–80% confluency (cell type dependent)
Number of Passages (cell type dependent)
DNA Quality and Quantity
The transfection protocol online library contains protocols obtained from the literature, developed by Bio-Rad scientists, or submitted by scientists like you. Browse protocols to view our library and find your starting point or submit a protocol by clicking the proper technology.
Belyansteva IA (2009). Helios Gene Gun-mediated transfection of the inner ear sensory epithelium. Methods Mol Biol 493, 103–123. PMID: 18839344
Fujiki R et al. (2009). GlcNAcylation of a histone methyltransferase in retinoic-acid-induced granulopoiesis. Nature 459, 455–459. PMID: 19377461
Helledie T et al. (2008). A simple and reliable electroporation method for human bone marrow mesenchymal stem cells. Stem Cells Dev 17, 837–848. PMID: 18752428
Hockemeyer D et al. (2009). Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases. Nat Biotechnol 27, 851–857. PMID: 19680244
Huang B et al. (2008). RNA interference-mediated in vivo silencing of fas ligand as a strategy for the enhancement of DNA vaccine potency. Hum Gene Ther 19, 763–773. PMID: 18627219
Shimamura K et al. (2007). Generation of secondary small interfering RNA in cell-autonomous and non-cell autonomous RNA silencing in tobacco. Plant Mol Biol 63, 803–813. PMID: 17225952
Su L et al. (2009). Neural stem cell differentiation is mediated by integrin beta4 in vitro. Int J Biochem Cell Biol 41, 916–924. PMID: 18834954
Tseng CN et al. (2013). A method to identify RNA A-to-I editing targets using I-specific cleavage and exon array analysis. Mol Cell Probes 7, 38–45. PMID: 22960667
Call us at 1-800-4-BIORAD (1-800-424-6723)
Please reenter your email address in the correct format.
Please enter your email address.
Your subscription information already exists, we will send you an email with specific instructions to manage your existing subscription profile.
To receive the latest news, promotions, and more, sign up for Bio-Rad updates by entering your email address below. You can elect to receive only the types of Bio-Rad communications that are of interest to you..
Copyright © 2014 Bio-Rad Laboratories, Inc. All rights reserved.