Interrogation of Cellular Innate Immunity by Diamond-Nanoneedle-Assisted Intracellular Molecular Fishing

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalNot applicablepeer-review

11 Scopus Citations
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Detail(s)

Original languageEnglish
Pages (from-to)7058-7063
Journal / PublicationNano Letters
Volume15
Issue number10
Early online date11 Sep 2015
Publication statusPublished - 14 Oct 2015

Abstract

Understanding intracellular signaling cascades and network is one of the core topics in modern biology. Novel tools based on nanotechnologies have enabled probing and analyzing intracellular signaling with unprecedented sensitivity and specificity. In this study, we developed a minimally invasive method for in situ probing specific signaling components of cellular innate immunity in living cells. The technique was based on diamond-nanoneedle arrays functionalized with aptamer-based molecular sensors, which were inserted into cytoplasmic domain using a centrifugation controlled process to capture molecular targets. Simultaneously, these diamond-nanoneedles also facilitated the delivery of double-strand DNAs (dsDNA90) into cells to activate the pathway involving the stimulator of interferon genes (STING). We showed that the nanoneedle-based biosensors can be successfully utilized to isolate transcriptional factor, NF-κB, from intracellular regions without damaging the cells, upon STING activation. By using a reversible protocol and repeated probing in living cells, we were able to examine the singling dynamics of NF-κB, which was quickly translocated from cytoplasm to nucleus region within 40 min of intracellular introduction of dsDNA90 for both A549 and neuron cells. These results demonstrated a novel and versatile tool for targeted in situ dissection of intracellular signaling, providing the potential to resolve new sights into various cellular processes.

Research Area(s)

  • Diamond nanoneedles, in situ detection, innate immunity, intracellular sensing, STING activation