Plug-and-play protein biosensors using aptamer-regulated in vitro transcription

Heonjoon Lee, Tian Xie, Byunghwa Kang, Xinjie Yu, Samuel W. Schaffter () and Rebecca Schulman ()
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Heonjoon Lee: Johns Hopkins University School of Medicine
Tian Xie: Johns Hopkins Bloomberg School of Public Health
Byunghwa Kang: Johns Hopkins University
Xinjie Yu: Johns Hopkins University
Samuel W. Schaffter: National Institute of Standards and Technology
Rebecca Schulman: Johns Hopkins University

Nature Communications, 2024, vol. 15, issue 1, 1-11

Abstract: Abstract Molecular biosensors that accurately measure protein concentrations without external equipment are critical for solving numerous problems in diagnostics and therapeutics. Modularly transducing the binding of protein antibodies, protein switches or aptamers into a useful output remains challenging. Here, we develop a biosensing platform based on aptamer-regulated transcription in which aptamers integrated into transcription templates serve as inputs to molecular circuits that can be programmed to a produce a variety of responses. We modularly design molecular biosensors using this platform by swapping aptamer domains for specific proteins and downstream domains that encode different RNA transcripts. By coupling aptamer-regulated transcription with diverse transduction circuits, we rapidly construct analog protein biosensors and digital protein biosensors with detection ranges that can be tuned over two orders of magnitude and can exceed the binding affinity of the aptamer. Aptamer-regulated transcription is a straightforward and inexpensive approach for constructing programmable protein biosensors that could have diverse applications in research and biotechnology.

Date: 2024
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DOI: 10.1038/s41467-024-51907-4

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