Synthesis of length-tunable DNA carriers for nanopore sensing

Molecular carriers represent an increasingly common strategy in the field of nanopore sensing to use secondary molecules to selectively report on the presence of target analytes in solution, allowing for sensitive assays of otherwise hard-to-detect molecules such as small, weakly-charged proteins. H...

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Bibliographic Details
Published in:PloS one 2023-08, Vol.18 (8), p.e0290559-e0290559
Main Authors: Roelen, Zachary, Tabard-Cossa, Vincent
Format: Article
Language:English
Subjects:
Analysis
Biology and Life Sciences
Biosensors
Chemical synthesis
Deoxyribonucleic acid
Design
DNA
DNA biosynthesis
Engineering and Technology
Enzymes
Genetic research
Genomes
Health aspects
Multiplexing
Nanotechnology
Physical sciences
Research and Analysis Methods
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Summary:Molecular carriers represent an increasingly common strategy in the field of nanopore sensing to use secondary molecules to selectively report on the presence of target analytes in solution, allowing for sensitive assays of otherwise hard-to-detect molecules such as small, weakly-charged proteins. However, existing carrier designs can often introduce drawbacks to nanopore experiments including higher levels of cost/complexity and carrier-pore interactions that lead to ambiguous signals and elevated clogging rates. In this work, we present a simple method of carrier production based on sticky-ended DNA molecules that emphasizes ease-of-synthesis and compatibility with nanopore sensing and analysis. In particular, our method incorporates the ability to flexibly control the length of the DNA carriers produced, enhancing the multiplexing potential of this carrier system through the separable nanopore signals they could generate for distinct targets. A proof-of-concept nanopore experiment is also presented, involving carriers produced by our method with multiple lengths and attached to DNA nanostructure targets, in order to validate the capabilities of the system. As the breadth of applications for nanopore sensors continues to expand, the availability of tools such as those presented here to help translate the outcomes of these applications into robust nanopore signals will be of major importance.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0290559