MolecularCloud January Newsletter

Electrode-free nanopore sensing by DiffusiOptoPhysiology

Dr. Shuo Huang's team published the paper "Electrode-free nanopore sensing by DiffusiOptoPhysiology" at ScienceAdvances in 06 Sep 2019. The plasmids of this article have been deposited on MolecularCloud.

A wide variety of single molecules can be identified by nanopore sensing. However, all reported nanopore sensing applications result from the same measurement configuration adapted from electrophysiology. Although urgently needed in commercial nanopore sequencing, parallel electrophysiology recording is limited in its cost and its throughput due to the introduced complexities from electronic integration.

Dr. Shuo Huang's team present the first electrode-free nanopore sensing method defined as DiffusiOptoPhysiology (DOP), in which single-molecule events are monitored optically without any electrical connections. Single-molecule sensing of small molecules, macromolecules, and biomacromolecules was subsequently demonstrated. As a further extension, a fingertip-sized, multiplexed chip with single-molecule sensing capabilities has been introduced, which suggests a new concept of clinical diagnosis using disposable nanopore sensors. DOP, which is universally compatible with all types of channels and a variety of fluorescence imaging platforms, may benefit diverse areas such as nanoporesequencing, drug screening, and channel protein investigations.

Plasmids from this article in MolecularCloud

Cat. No.

Plasmid Name

Description

MC_0068416

α-HL WT D8H6

The plasmid α-HL WT D8H6 is designed for the preparation of the heptameric transmembrane protein alpha-hemolysin (α-HL) WT by prokaryotic expression. A H6 tag is designed for the followup purification using nickel affinity chromatography. α-HL is naturally secreted by Staphylococcus aureus. The structure has been characterized using X-ray crystallography (Song L, Hobaugh M R, Shustak C, et al. Structure of staphylococcal α-hemolysin, a heptameric transmembrane pore. Science, 1996, 274(5294): 1859-1865.). Since 1996, α-HL WT along with its mutants have been widely investigated as single-molecule nanopore sensors, suitable for sensing a variety of analyte such as ionic species, small molecules, macromolecules and biomacromolecules.

MC_0068760

ClyA RR

The plasmid ClyA-RR is designed for the preparation of a mutant Cytolysin A (ClyA). ClyA is a pore-forming toxin which was found in various E. coli strains. Its structure has been characterized using X-ray crystallography (PDB: 2WCD).With a unique wide aperture, ClyA along with its mutants have been used as nanopore sensors for the detection of macromolecules. The ClyA-RR mutant is engineered for more efficient DNA sensing (Franceschini L, Brouns T, Willems K. et al. DNA Translocation through Nanopores at Physiological Ionic Strengths Requires Precise Nanoscale Engineering. ACS Nano 2016, 10(9): 8394-8402). It is also a highly compatible sensor for the imaging based DiffusiOptoPhysiology platform (Wang Y, Wang Y, Du X, et al. Electrode-free nanopore sensing by DiffusiOptoPhysiology[J]. Science Advances, 2019, 5(9): eaar3309.)


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