Samenvatting
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.
Originele taal-2 | Engels |
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Pagina's (van-tot) | 18546-18555 |
Aantal pagina's | 10 |
Tijdschrift | Angewandte Chemie - International Edition |
Volume | 59 |
Nummer van het tijdschrift | 42 |
Vroegere onlinedatum | 6 jul. 2020 |
DOI's | |
Status | Gepubliceerd - 12 okt. 2020 |
Financiering
This work was financially supported by the Netherlands Science Organization (Grant 192.028) through the VIDI program, the Ministerio de Economía Competitividad/Agencia Estatal de Investigación (AEI) (EURONANOMED/Acciones de Programación Conjunta Internacional PCIN‐2016‐025), the European Union's Horizon 2020 research and innovation program under the European Research Council grant ERC‐StG‐757397 (NANOSTORM) and the Marie Sklodowska‐Curie grant agreement 765497 (THERACAT), the “La Caixa” Foundation, the Generalitat de Catalunya (2017 SGR 01536), the MU Life Sciences Center (LSC) ‐ Early Concept Grant (ECG) 2019 for Innovative Collaborative Research involving Post‐Doctoral Researchers, and the UM Research and Creative Works Strategic Investment Program grant. y This work was financially supported by the Netherlands Science Organization (Grant 192.028) through the VIDI program, the Ministerio de Econom?a y Competitividad/Agencia Estatal de Investigaci?n (AEI) (EURONANOMED/Acciones de Programaci?n Conjunta Internacional PCIN-2016-025), the European Union's Horizon 2020 research and innovation program under the European Research Council grant ERC-StG-757397 (NANOSTORM) and the Marie Sklodowska-Curie grant agreement 765497 (THERACAT), the ?La Caixa? Foundation, the Generalitat de Catalunya (2017 SGR 01536), the MU Life Sciences Center (LSC) - Early Concept Grant (ECG) 2019 for Innovative Collaborative Research involving Post-Doctoral Researchers, and the UM Research and Creative Works Strategic Investment Program grant.
Financiers | Financiernummer |
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H2020 Marie Skłodowska-Curie Actions | |
European Union’s Horizon Europe research and innovation programme | 757397 |
European Research Council | 765497 |
Generalitat de Catalunya | 2017 SGR 01536 |