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Design of Split Proximity Circuit as a Plug-and-Play Translator for Point Mutation Discrimination.

Y. S. Ang,P. Lai,Lin‐Yue Lanry Yung

Published 2020 in Analytical Chemistry

ABSTRACT

Point mutations are the a common form of genetic variation and have been identified as important disease biomarkers. Conventional methods for analyzing point mutations, e.g. PCR, are based on differences in thermal stability of the DNA duplex, which require extensive optimization of the reaction condition and non-trivial design of sequence-selective primers. This motivated the design of molecular translators to convert molecular inputs into generic output sequences which allows for the target recognition and signal generation regions to be designed independently. In this work, we propose a translator design based on the concept of split proximity circuit (SPC) to achieve both high sequence selectivity and assay robustness using a universal reaction condition, i.e. room temperature and constant ionic concentration. We discussed the design aspects of the SPC recognition regions and demonstrated its plug-and-play capability to discriminate different point mutations for both DNA (seven G6PD mutations) and RNA (let-7 microRNA family members) targets while retaining the same signal generation region. Despite its simple design and non-stringent assay condition requirements, the SPC retained good analytical performance to detect sub-nanomolar target concentration within a reasonable time of an hour.

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