PLANT-Dx: A Molecular Diagnostic for Point-of-Use Detection of Plant Pathogens

ACS Synthetic Biology
M VerosloffJ B Lucks

Abstract

Synthetic biology based diagnostic technologies have improved upon gold standard diagnostic methodologies by decreasing cost, increasing accuracy, and enhancing portability. However, there has been little effort in adapting these technologies toward applications related to point-of-use monitoring of plant and crop health. Here, we take a step toward this vision by developing an approach that couples isothermal amplification of specific plant pathogen genomic sequences with customizable synthetic RNA regulators that are designed to trigger the production of a colorimetric output in cell-free gene expression reactions. We demonstrate our system can sense viral derived sequences with high sensitivity and specificity, and can be utilized to directly detect viruses from infected plant material. Furthermore, we demonstrate that the entire system can operate using only body heat and naked-eye visual analysis of outputs. We anticipate these strategies to be important components of user-friendly and deployable diagnostic systems that can be configured to detect a range of important plant pathogens.

References

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Jun 8, 2006·PLoS Biology·Olaf PiepenburgNiall A Armes
Jul 21, 2010·Journal of Computational Chemistry·Joseph N ZadehNiles A Pierce
Oct 25, 2011·Molecular Plant Pathology·Karen-Beth G ScholthofGary D Foster
Mar 20, 2014·Molecular BioSystems·Lei YanHerman O Sintim
Dec 9, 2014·Frontiers in Plant Science·Valérie Nicaise
Feb 3, 2015·Nature Chemical Biology·James ChappellJulius B Lucks
Nov 26, 2015·Proceedings of the National Academy of Sciences of the United States of America·Shimyn SlomovicJames J Collins
Jan 29, 2016·ACS Synthetic Biology·Jonathan GaramellaVincent Noireaux
Apr 15, 2017·Science·Jonathan S GootenbergFeng Zhang
Nov 16, 2018·Biochemistry·Paul D Carlson, Julius B Lucks

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Citations

Nov 30, 2019·Nature Reviews. Genetics·Adam D SilvermanMichael C Jewett
Feb 3, 2021·Nature Communications·Evan AmalfitanoKeith Pardee
Feb 27, 2021·Frontiers in Microbiology·Ilenne Del ValleLauren B Stadler
Jun 12, 2020·ACS Synthetic Biology·Weston KightlingerMichael C Jewett
Jan 4, 2021·Trends in Biotechnology·Margrethe A Boyd, Neha P Kamat
Apr 27, 2021·Annual Review of Chemical and Biomolecular Engineering·Walter ThavarajahJulius B Lucks
May 22, 2021·Trends in Genetics : TIG·Sneha GulatiDebojyoti Chakraborty
Jun 8, 2021·Frontiers in Bioengineering and Biotechnology·Xinjie Chen, Yuan Lu
Aug 7, 2021·Frontiers in Bioengineering and Biotechnology·Megan A McSweeney, Mark P Styczynski
Dec 12, 2019·ACS Synthetic Biology·Xiangyang LiuSrivatsan Raman
Dec 13, 2019·ACS Synthetic Biology·Walter ThavarajahJulius B Lucks
Feb 23, 2020·ACS Synthetic Biology·Adam D SilvermanJulius B Lucks
Sep 10, 2021·Frontiers in Bioengineering and Biotechnology·Anibal ArceFernán Federici
Sep 24, 2021·Advanced Materials·Simone GiaveriFrancesco Stellacci
Oct 1, 2021·Nature Communications·Yan ZhangMark P Styczynski

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Methods Mentioned

BETA
in vitro transcription

Software Mentioned

NUPACK
PLANT

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