CRISPR-Mediated Strand Displacement Logic Circuits with Toehold-Free DNA.

María Heras-Hernández,Roser Montagud-Martínez,José-Antonio Daròs,Lucas Goiriz,Guillermo Rodrigo

doi:10.1021/acssynbio.0c00649

María Heras-Hernández, Roser Montagud-Martínez + Show 3 more

Open Access

https://doi.org/10.1021/acssynbio.0c00649

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Abstract

DNA nanotechnology, and DNA computing in particular, has grown extensively over the past decade to end with a variety of functional stable structures and dynamic circuits. However, the use as designer elements of regular DNA pieces, perfectly complementary double strands, has remained elusive. Here, we report the exploitation of CRISPR-Cas systems to engineer logic circuits based on isothermal strand displacement that perform with toehold-free double-stranded DNA. We designed and implemented molecular converters for signal detection and amplification, showing good interoperability between enzymatic and nonenzymatic processes. Overall, these results contribute to enlarge the repertoire of substrates and reactions (hardware) for DNA computing.

Highlights

deoxyribonucleic acid (DNA) nanotechnology, and DNA computing in particular, has grown extensively over the past decade to end with a variety of functional stable structures and dynamic circuits
We introduce the concept of CRISPR-mediated strand displacement in order to work with regular dsDNA in logic circuits (CRISPR stands for clustered regularly interspaced short palindromic repeats);[14] that is, to exploit as functional elements, rather than being mere waste products, dsDNA molecules that lack toeholds
We proved the suitability of this approach by engineering different logic circuits responsive to toehold-free dsDNA molecules, producing as outputs individual oligonucleotides

Summary

Introduction

DNA nanotechnology, and DNA computing in particular, has grown extensively over the past decade to end with a variety of functional stable structures and dynamic circuits. Molecule that is produced in a conventional toehold-mediated strand displacement reaction as an intermediate species thanks to a given sgRNA and Cas9n (Figure 2b).

Results

Conclusion