Aptamers with Tunable Affinity Enable Single-Molecule Tracking and Localization of Membrane Receptors on Living Cancer Cells

Pietro Delcanale; David Porciani; Silvia Pujals; Alexander Jurkevich; Andrian Chetrusca; Kwaku D. Tawiah; Donald H. Burke; Lorenzo Albertazzi

doi:10.1002/anie.202004764

Aptamers with Tunable Affinity Enable Single-Molecule Tracking and Localization of Membrane Receptors on Living Cancer Cells

Pietro Delcanale, David Porciani, Silvia Pujals, Alexander Jurkevich, Andrian Chetrusca, Kwaku D. Tawiah, Donald H. Burke (Corresponding author), Lorenzo Albertazzi (Corresponding author)

Research output: Contribution to journal › Article › Academic › peer-review

51 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	18546-18555
Number of pages	10
Journal	Angewandte Chemie - International Edition
Volume	59
Issue number	42
Early online date	6 Jul 2020
DOIs	https://doi.org/10.1002/anie.202004764
Publication status	Published - 12 Oct 2020

Funding

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.

Funders	Funder number
Marie Skłodowska‐Curie
European Union's Horizon 2020 - Research and Innovation Framework Programme	757397
H2020 European Research Council	765497
Generalitat de Catalunya	2017 SGR 01536

Keywords

aptamers
cell-surface receptors
live-cell imaging
PAINT
single-molecule tracking

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1002/anie.202004764

Cite this

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abstract = "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.",

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AU - Chetrusca, Andrian

AU - Tawiah, Kwaku D.

AU - Burke, Donald H.

AU - Albertazzi, Lorenzo

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Aptamers with Tunable Affinity Enable Single-Molecule Tracking and Localization of Membrane Receptors on Living Cancer Cells

Abstract

Funding

Keywords

UN SDGs

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