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Revision as of 04:05, 14 September 2015

Abstract

In the past year, paper­based transcription and translation, reconstituted from freeze­drying, have been adapted in a variety of ways and shown to be effective after a year of storage at room temperature. However, currently, this technology is severely limited in its applications because protein detection requires a different strategy for each individual protein. Aptapaper uses the targeting specificity of aptamers to create a modular protein detection system that can easily be adapted to any protein. A DNA aptamer is bound to a DNA trigger and becomes unbound in response to its protein of specificity. This frees the DNA trigger to turn on an RNA toehold switch, resulting in a 40 fold change in reporter protein expression, with more results to come. This system, when freeze­dried on paper, is cheap and portable, making it well suited to tackle the unmet needs for disease detection in remote areas.

Aptapaper uses a genetic switch system and the targeting specificity of aptamers to detect virtually any protein, all on a simple piece of filter paper. A DNA aptamer is bound to a DNA trigger and becomes unbound in response to its protein of specificity. This frees the DNA trigger to turn on an RNA toehold switch, resulting in translation of a reporter protein. This system can easily be freeze­dried on paper, creating a cheap, durable device. We used a toehold switch design that can easily be adapted to any trigger RNA, while still maintaining fold changes over 100, at very low background. The trigger can thus be adapted to any aptamer, and the modularity of these toehold switches is maintained.

Equally important, versatile toehold switches have been optimized and demonstrated to turn on a gene circuit in response to virtually any trigger RNA.