A Gold Nanoparticle Bio-Optical Transponder to Dynamically Monitor Intracellular pH

作者：Carnevale Kate J F; Riskowski Ryan A; Strouse Geoffrey F^*

来源：ACS Nano, 2018, 12(6): 5956-5968.

DOI：10.1021/acsnano.8b02200

• intracellular pH
• nanoprobe
• gold nanoparticle
• AuNP
• multicolor SET
• bio-optical transponder

A pH-sensitive bio-optical transponder (pH-BOT) capable of simultaneously reporting the timing of intracellular DNA cargo release from a gold nanoparticle (AuNP) and the evolving intracellular pH (pHi) during endosomal maturation is demonstrated. The pH-BOT is designed with a triple-dye-labeled duplex DNA appended to a 6.6 nm AuNP, utilizing pH-responsive fluorescein paired with DyLight405 as a surface energy transfer (SET) coupled dye pair to ratiometrically report the pH at and after cargo release. A non-SET-coupled dye, DyLight 700, is used to provide dynamic tracking throughout the experiment. The pH-BOT beacon of the cargo uptake, release, and processing was visualized using live-cell confocal fluorescent microscopy in Chinese hamster ovary cells, observed that while maturation of endosomes carrying pH-BOT is slowed significantly, the pH-BOT is throughout the endolysosomal system while remaining at pH similar to 6. This observed decoupling of endosomal maturation from acidification lends support to those models that propose that pH alone is not sufficient to explain endosomal maturation and may enable greater insight into our understanding of the fundamental processes of biology.

• 出版日期2018-6