Aptamer-based optical manipulation of protein subcellular localization in cells

Abstract

Protein-dominant cellular processes cannot be fully decoded without precise manipulation of their activity and localization in living cells. Advances in optogenetics have allowed spatiotemporal control over cellular proteins with molecular specificity; however, these methods require recombinant expression of fusion proteins, possibly leading to conflicting results. Instead of modifying proteins of interest, in this work, we focus on design of a tunable recognition unit and develop an aptamer-based near-infrared (NIR) light-responsive nanoplatform for manipulating the subcellular localization of specific proteins in their native states. Our results demonstrate that this nanoplatform allows photocontrol over the cytoplasmicnuclear shuttling behavior of the target RelA protein (a member of the NF-κβ family), enabling regulation of RelA-related signaling pathways. With a modular design, this aptamer-based nanoplatform can be readily extended for the manipulation of different proteins (e.g., lysozyme and p53), holding great potential to develop a variety of label-free protein photoregulation strategies for studying complex biological events.

An Overview on SARS-CoV-2 (COVID-19) and Other Human Coronaviruses and Their Detection Capability via Amplification Assay, Chemical Sensing, Biosensing, Immunosensing, and Clinical Assays

“…Another interesting feature of aptamers is that they effectively preserve their properties in various experimental conditions [ 149 , 150 ]. Aptasensing bio-assays have various recognition mechanisms and can bind to a wide range of regulatory proteins [ 138 , 151 ], enzymes [ 152 , 153 ], mono- and polyclonal antibodies [ 154 ], amino acids [ 155 ], growth factors [ 156 ], toxins [ 157 ], low molecular weight vitamins [ 158 ], cancer biomarkers [ 159 – 161 ], and even some metal ions [ 162 ]. Large quantities of aptamers can be biochemically synthesized most frequently through the systematic evolution of ligands by exponential enrichment (SELEX).…”

Peptide and protein assays using customizable bio-affinity arrays combined with ambient ionization mass spectrometry

“…Nucleic acid aptamer can be regarded as an emerging artificial antibody that can bind to a specific target molecule while possessing higher synthetic availability and stability than antibody. [60][61][62] We switched the bio-affinity materials to thrombin binding aptamer-grafted gold nanoparticles (TBA-AuNPs) (Fig. 1e), and constructed a "proof-of-concept" MS assaying platform of protein biomarkers by ambient ionization (see Supporting Information for the detailed experimental procedures).…”

Advanced Optogenetic-Based Biosensing and Related Biomaterials

“…Optical tools that modulate protein-protein interactions emerged: opening the door to optogenetic control of kinases and transcription factors. The design of a tunable recognition unit and development of an aptamer-based near-infrared (NIR) light-responsive nanoplatform for manipulating the subcellular localization of specific proteins in their native states was reported [ 35 ].…”