Debugging Eukaryotic Genetic Code Expansion for Site‐Specific Click‐PAINT Super‐Resolution Microscopy
Abstract: Super‐resolution microscopy (SRM) greatly benefits from the ability to install small photostable fluorescent labels into proteins. Genetic code expansion (GCE) technology addresses this demand, allowing the introduction of small labeling sites, in the form of uniquely reactive noncanonical amino acids (ncAAs), at any residue in a target protein. However, low incorporation efficiency of ncAAs and high background fluorescence limit its current SRM applications. Redirecting the subcellular localization of the pyrrolysine‐based GCE system for click chemistry, combined with DNA‐PAINT microscopy, enables the visualization of even low‐abundance proteins inside mammalian cells. This approach links a versatile, biocompatible, and potentially unbleachable labeling method with residue‐specific precision. Moreover, our reengineered GCE system eliminates untargeted background fluorescence and substantially boosts the expression yield, which is of general interest for enhanced protein engineering in eukaryotes using GCE.
- Location
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Deutsche Nationalbibliothek Frankfurt am Main
- Extent
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Online-Ressource
- Language
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Englisch
- Bibliographic citation
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Debugging Eukaryotic Genetic Code Expansion for Site‐Specific Click‐PAINT Super‐Resolution Microscopy ; volume:55 ; number:52 ; year:2016 ; pages:16172-16176 ; extent:5
Angewandte Chemie / International edition. International edition ; 55, Heft 52 (2016), 16172-16176 (gesamt 5)
- Creator
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Nikić-Spiegel, Ivana
Estrada Girona, Gemma
Kang, Jun Hee
Paci, Giulia
Mikhaleva, Sofya
Koehler, Christine
Shymanska, Nataliia V.
Ventura Santos, Camilla
Spitz, Daniel
Lemke, Edward A.
- DOI
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10.1002/anie.201608284
- URN
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urn:nbn:de:101:1-2022101406030952393522
- Rights
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Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
- Last update
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15.08.2025, 7:21 AM CEST
Data provider
Deutsche Nationalbibliothek. If you have any questions about the object, please contact the data provider.
Associated
- Nikić-Spiegel, Ivana
- Estrada Girona, Gemma
- Kang, Jun Hee
- Paci, Giulia
- Mikhaleva, Sofya
- Koehler, Christine
- Shymanska, Nataliia V.
- Ventura Santos, Camilla
- Spitz, Daniel
- Lemke, Edward A.
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