Abstract
SNAP display is based on the covalent reaction of the DNA repair protein AGT (O6-alkylguanine DNA alkyltransferase, the “SNAP-tag”) with its substrate benzylguanine (BG). Linear, BG-labelled template DNA is encapsulated in water-in-oil emulsion droplets with a diameter of a few micrometres (i.e. 1 mL of emulsion contains ∼1010 compartments). Each droplet contains only a single DNA copy, which is transcribed and translated in vitro. The expressed AGT fusion proteins attach to their coding DNA via the BG label inside the droplet, which ensures that a specific genotype–phenotype linkage is established. Subsequently, the emulsion is broken and protein-DNA conjugates, which constitute a DNA-tagged protein library, selected via affinity panning. This method will prove a useful addition to the array of in vitro display systems, distinguished by the stability of DNA as the coding nucleic acid and the covalent link between gene and protein.
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Notes
- 1.
The SNAP-tag technology was commercialised by Covalys AG in 2007. Since 2009, it is distributed by New England Biolabs.
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Acknowledgements
MK was supported by a fellowship from the EU Marie-Curie ITN ProSA. FH is an ERC Starting Investigator.
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Kaltenbach, M., Hollfelder, F. (2012). SNAP Display: In Vitro Protein Evolution in Microdroplets. In: Douthwaite, J., Jackson, R. (eds) Ribosome Display and Related Technologies. Methods in Molecular Biology, vol 805. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-379-0_7
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