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A simple method for spray-on gene editing in planta

Cara Doyle, View ORCID ProfileKatie Higginbottom, View ORCID ProfileThomas A. Swift, View ORCID ProfileMark Winfield, View ORCID ProfileChristopher Bellas, David Benito-Alifonso, Taryn Fletcher, View ORCID ProfileM. Carmen Galan, Keith Edwards, View ORCID ProfileHeather M. Whitney
doi: https://doi.org/10.1101/805036
Cara Doyle
1School of Biological Sciences, Life Sciences Building, University of Bristol, BS8 1TQ, UK2
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Katie Higginbottom
1School of Biological Sciences, Life Sciences Building, University of Bristol, BS8 1TQ, UK2
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  • ORCID record for Katie Higginbottom
Thomas A. Swift
2School of Chemistry, University of Bristol, BS8 1TS, UK
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Mark Winfield
1School of Biological Sciences, Life Sciences Building, University of Bristol, BS8 1TQ, UK2
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  • ORCID record for Mark Winfield
Christopher Bellas
1School of Biological Sciences, Life Sciences Building, University of Bristol, BS8 1TQ, UK2
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  • ORCID record for Christopher Bellas
David Benito-Alifonso
2School of Chemistry, University of Bristol, BS8 1TS, UK
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Taryn Fletcher
1School of Biological Sciences, Life Sciences Building, University of Bristol, BS8 1TQ, UK2
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M. Carmen Galan
2School of Chemistry, University of Bristol, BS8 1TS, UK
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  • ORCID record for M. Carmen Galan
  • For correspondence: heather.whitney@bristol.ac.uk
Keith Edwards
1School of Biological Sciences, Life Sciences Building, University of Bristol, BS8 1TQ, UK2
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  • For correspondence: heather.whitney@bristol.ac.uk
Heather M. Whitney
1School of Biological Sciences, Life Sciences Building, University of Bristol, BS8 1TQ, UK2
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  • ORCID record for Heather M. Whitney
  • For correspondence: heather.whitney@bristol.ac.uk
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Abstract

Potential innovation in Plant research through the use of gene-edited and genetically modified plants is currently being hindered by inefficient and costly plant transformation. We show that naturally occurring carbon dots (quasi-spherical, <10nm nanoparticles) can act as a fast vehicle for carrying plasmids into mature plant cells, resulting in transient plant transformation in a number of important crop species with no negative impacts on photosynthesis or growth. We further show that GFP, Cas9, and gRNA introduced into wheat via foliar application (spraying on) of plasmid coated carbon dots are expressed and, in the case of Cas9, make genome edits in SPO11 genes. Therefore, we present a protocol for spray-on gene editing that is simple, inexpensive, fast, transforms in planta, and is applicable to multiple crop species. We believe this technique creates many opportunities for the future of plant transformation in research and shows great promise for plant protein production systems.

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Posted October 25, 2019.
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A simple method for spray-on gene editing in planta
Cara Doyle, Katie Higginbottom, Thomas A. Swift, Mark Winfield, Christopher Bellas, David Benito-Alifonso, Taryn Fletcher, M. Carmen Galan, Keith Edwards, Heather M. Whitney
bioRxiv 805036; doi: https://doi.org/10.1101/805036
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A simple method for spray-on gene editing in planta
Cara Doyle, Katie Higginbottom, Thomas A. Swift, Mark Winfield, Christopher Bellas, David Benito-Alifonso, Taryn Fletcher, M. Carmen Galan, Keith Edwards, Heather M. Whitney
bioRxiv 805036; doi: https://doi.org/10.1101/805036

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