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Aptamers with Tunable Affinity Enable Single‐Molecule Tracking and Localization of Membrane Receptors on Living Cancer Cells
Author(s) -
Delcanale Pietro,
Porciani David,
Pujals Silvia,
Jurkevich Alexander,
Chetrusca Andrian,
Tawiah Kwaku D.,
Burke Donald H.,
Albertazzi Lorenzo
Publication year - 2020
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.202004764
Subject(s) - aptamer , receptor , biophysics , cell , microbiology and biotechnology , cell surface receptor , cell membrane , fluorescence , cancer cell , fluorescence lifetime imaging microscopy , tracking (education) , chemistry , molecular imaging , computational biology , biology , cancer , biochemistry , genetics , physics , optics , in vivo , psychology , pedagogy
Tumor cell‐surface markers are usually overexpressed or mutated protein receptors for which spatiotemporal regulation differs between and within cancers. Single‐molecule fluorescence imaging can profile individual markers in different cellular contexts with molecular precision. However, standard single‐molecule imaging methods based on overexpressed genetically encoded tags or cumbersome probes can significantly alter the native state of receptors. We introduce a live‐cell points accumulation for imaging in nanoscale topography (PAINT) method that exploits aptamers as minimally invasive affinity probes. Localization and tracking of individual receptors are based on stochastic and transient binding between aptamers and their targets. We demonstrated single‐molecule imaging of a model tumor marker (EGFR) on a panel of living cancer cells. Affinity to EGFR was finely tuned by rational engineering of aptamer sequences to define receptor motion and/or native receptor density.