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Direct Comparison of d-DNA and l-DNA Strand-Displacement Reactions in Living Mammalian Cells
Author(s) -
Wenrui Zhong,
Jonathan T. Sczepanski
Publication year - 2020
Publication title -
acs synthetic biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.156
H-Index - 66
ISSN - 2161-5063
DOI - 10.1021/acssynbio.0c00527
Subject(s) - dna , multiple displacement amplification , biophysics , dna damage , chemistry , biology , microbiology and biotechnology , biochemistry , polymerase chain reaction , gene , dna extraction
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.

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