caa a22 4500 605479291 CHVBK 20210128100409.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s10544-014-9908-7 doi (NATIONALLICENCE)springer-10.1007/s10544-014-9908-7 Detection of GNAQ mutations and reduction of cell viability in uveal melanoma cells with functionalized gold nanoparticles [Elektronische Daten] [Christian Posch, Alfonso Latorre, Michelle Crosby, Anna Celli, Ana Latorre, Igor Vujic, Martina Sanlorenzo, Gary Green, Jingly Weier, Mitchell Zekhtser, Jeffrey Ma, Gabriela Monico, Devron Char, Denis Jusufbegovic, Klemens Rappersberger, Álvaro Somoza, Susana Ortiz-Urda] Background: Uveal melanoma (UM) is the most common primary intraocular malignancy in adults. Early treatment may improve any chances of preventing metastatic disease, but diagnosis of small UM is challenging. Up to 95% of all UMs carry somatic mutations in the G-coupled proteins GNAQ and GNA11 promoting anchorage-independent growth and proliferation. About 50% of UMs are fatal. Once metastatic, patients have limited options for successful therapy. Methods: We have developed functionalized gold nanoparticles (AuNPs) to visualize transcripts of mutant GNAQ mRNA in living cells. In addition to their suitability as a specific tool for GNAQ mutation detection, we have developed a novel linker that enables conjugation of siRNAs to AuNPs allowing for greater and more rapid intracellular release of siRNAs compared to previously described approaches. Results: Binding of modified AuNPs to matching target mRNA leads to conformational changes, resulting in a detectable fluorescent signal that can be used for mutation detection in living cells. Knockdown of GNAQ with siRNA-AuNPs effectively reduced downstream signals and decreased cell viability in GNAQ mutant uveal melanoma cells. Conclusion: AuNPs may in future be developed to serve as sensors for mutations of vital importance. The new release system for siRNA-AuNP improves previous systems, which conceivably will be useful for future therapeutic gene regulatory approaches. Springer Science+Business Media New York, 2015 siRNA nationallicence GNAQ nationallicence Fluorescence nationallicence Treatment nationallicence Linker nationallicence Posch Christian Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut Latorre Alfonso Instituto Madrileño de Estudios Avanzados en Nanociencia (IMDEA Nanociencia), & CNB-CSIC-IMDEA Nanociencia Associated Unit "Unidad de Nanobiotecnología”, 28049, Madrid, Spain aut Crosby Michelle North County Eye Surgery Associates, 320 Santa Fe Drive Suite 104, 92024, Encinitas, USA aut Celli Anna Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut Latorre Ana Instituto Madrileño de Estudios Avanzados en Nanociencia (IMDEA Nanociencia), & CNB-CSIC-IMDEA Nanociencia Associated Unit "Unidad de Nanobiotecnología”, 28049, Madrid, Spain aut Vujic Igor Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut Sanlorenzo Martina Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut Green Gary Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut Weier Jingly Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut Zekhtser Mitchell Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut Ma Jeffrey Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut Monico Gabriela Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut Char Devron Departments of Ophthalmology, Stanford University, University of California San Francisco and California Pacific Medical Center, 45 Castro Street, 94114, San Francisco, USA aut Jusufbegovic Denis Departments of Ophthalmology, Stanford University, University of California San Francisco and California Pacific Medical Center, 45 Castro Street, 94114, San Francisco, USA aut Rappersberger Klemens Department of Dermatology, The Rudolfstiftung Hospital, Juchgasse 25, 1030, Vienna, Austria aut Somoza Álvaro Department of Dermatology, The Rudolfstiftung Hospital, Juchgasse 25, 1030, Vienna, Austria aut Ortiz-Urda Susana Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut Biomedical Microdevices Springer US; http://www.springer-ny.com 17/1(2015-02-01), 1-9 1387-2176 17:1<1 2015 17 10544 https://doi.org/10.1007/s10544-014-9908-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10544-014-9908-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Posch Christian Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut NATIONALLICENCE 700 1- Latorre Alfonso Instituto Madrileño de Estudios Avanzados en Nanociencia (IMDEA Nanociencia), & CNB-CSIC-IMDEA Nanociencia Associated Unit "Unidad de Nanobiotecnología”, 28049, Madrid, Spain aut NATIONALLICENCE 700 1- Crosby Michelle North County Eye Surgery Associates, 320 Santa Fe Drive Suite 104, 92024, Encinitas, USA aut NATIONALLICENCE 700 1- Celli Anna Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut NATIONALLICENCE 700 1- Latorre Ana Instituto Madrileño de Estudios Avanzados en Nanociencia (IMDEA Nanociencia), & CNB-CSIC-IMDEA Nanociencia Associated Unit "Unidad de Nanobiotecnología”, 28049, Madrid, Spain aut NATIONALLICENCE 700 1- Vujic Igor Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut NATIONALLICENCE 700 1- Sanlorenzo Martina Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut NATIONALLICENCE 700 1- Green Gary Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut NATIONALLICENCE 700 1- Weier Jingly Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut NATIONALLICENCE 700 1- Zekhtser Mitchell Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut NATIONALLICENCE 700 1- Ma Jeffrey Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut NATIONALLICENCE 700 1- Monico Gabriela Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut NATIONALLICENCE 700 1- Char Devron Departments of Ophthalmology, Stanford University, University of California San Francisco and California Pacific Medical Center, 45 Castro Street, 94114, San Francisco, USA aut NATIONALLICENCE 700 1- Jusufbegovic Denis Departments of Ophthalmology, Stanford University, University of California San Francisco and California Pacific Medical Center, 45 Castro Street, 94114, San Francisco, USA aut NATIONALLICENCE 700 1- Rappersberger Klemens Department of Dermatology, The Rudolfstiftung Hospital, Juchgasse 25, 1030, Vienna, Austria aut NATIONALLICENCE 700 1- Somoza Álvaro Department of Dermatology, The Rudolfstiftung Hospital, Juchgasse 25, 1030, Vienna, Austria aut NATIONALLICENCE 700 1- Ortiz-Urda Susana Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA aut NATIONALLICENCE 773 0- Biomedical Microdevices Springer US; http://www.springer-ny.com 17/1(2015-02-01), 1-9 1387-2176 17:1<1 2015 17 10544