Work

Direct Comparison of d -DNA and l -DNA Strand-Displacement Reactions in Living Mammalian Cells

Year: 2020

Type: article

Abstract: To overcome technical challenges associated with the use of DNA strand-displacement circuits in vivo, including degradation by cellular nucleases, researchers are increasingly turning to bio-orthogonal l -DNA. Although enhanced stability and improved performance of l -DNA-based circuits within living cells are often implied, direct experimental evidence has not been provided. Herein, we directly compare the functional stability and kinetics of d -DNA and l -DNA strand-displacement in live cells for the first time. We show that l -DNA strand-displacement reaction systems have minimal “leak”, fast reaction kinetics, and prolonged stability inside living cells as compared to conventional d -DNA. Furthermore, using “heterochiral” strand-displacement, we demonstrate that biostable l -DNA reaction components can be easily interfaced with native DNA inside cells. Overall, our results strongly support the broader adoption of l -DNA in the field of DNA molecular circuitry, especially for in vivo applications.

Source: ACS Synthetic Biology

DOI: 10.1021/acssynbio.0c00527

Authors: Wenrui Zhong, Jonathan T. Sczepanski

Institution: Texas A&M University

Language: English

Topic: Advanced biosensing and bioanalysis techniques

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: Cancer Prevention and Research Institute of Texas, National Institute of Biomedical Imaging and Bioengineering

Awards: Mirror image DNA circuitry for complex microRNA analysis in live cells, Recruitment of First-Time, Tenure-Track Faculty Members