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)

Onderzoeksoutput: Bijdrage aan tijdschrift › Tijdschriftartikel › Academic › peer review

53 Citaten (Scopus)

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
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	https://doi.org/10.1002/anie.202004764
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
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

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10.1002/anie.202004764

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anie.202004764
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title = "Aptamers with Tunable Affinity Enable Single-Molecule Tracking and Localization of Membrane Receptors on Living Cancer Cells",

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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Aptamers with Tunable Affinity Enable Single-Molecule Tracking and Localization of Membrane Receptors on Living Cancer Cells. / Delcanale, Pietro; Porciani, David; Pujals, Silvia et al.
In: Angewandte Chemie - International Edition, Vol. 59, Nr. 42, 12.10.2020, blz. 18546-18555.

Onderzoeksoutput: Bijdrage aan tijdschrift › Tijdschriftartikel › Academic › peer review

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AU - Porciani, David

AU - Pujals, Silvia

AU - Jurkevich, Alexander

AU - Chetrusca, Andrian

AU - Tawiah, Kwaku D.

AU - Burke, Donald H.

AU - Albertazzi, Lorenzo

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

Samenvatting

Financiering

Duurzame ontwikkelingsdoelstellingen van de VN

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