title: Detection of GNAQ mutations and reduction of cell viability in uveal melanoma cells with functionalized gold nanoparticles.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors: Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA, cposch81@gmail.com.
authors: Christian
authors: C
authors: Posch
authors: Alfonso
authors: A
authors: Latorre
authors: Michelle B
authors: MB
authors: Crosby
authors: Anna
authors: A
authors: Celli
authors: Ana
authors: A
authors: Latorre
authors: Igor
authors: I
authors: Vujic
authors: Martina
authors: M
authors: Sanlorenzo
authors: Gary A
authors: GA
authors: Green
authors: Jingly
authors: J
authors: Weier
authors: Mitchell
authors: M
authors: Zekhtser
authors: Jeffrey
authors: J
authors: Ma
authors: Gabriela
authors: G
authors: Monico
authors: Devron H
authors: DH
authors: Char
authors: Denis
authors: D
authors: Jusufbegovic
authors: Klemens
authors: K
authors: Rappersberger
authors: Álvaro
authors: Á
authors: Somoza
authors: Susana
authors: S
authors: Ortiz-Urda
mesh_terms: D000328:Adult; D015374:Biosensing Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics; D044385:GTP-Binding Protein alpha Subunits / Q000235:genetics / Q000378:metabolism; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D055785:Gene Knockdown Techniques / Q000379:methods*; D006046:Gold / Q000737:chemistry*; D006801:Humans; D008545:Melanoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D053768:Metal Nanoparticles / Q000737:chemistry*; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D014604:Uveal Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology
chemical_list: C554061:GNAQ protein, human; D044385:GTP-Binding Protein alpha Subunits; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D006046:Gold; D043802:GTP-Binding Protein alpha Subunits, Gq-G11
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