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A versatile CRISPR-based system for lineage tracing in living plants

View ORCID ProfileMattia Donà, View ORCID ProfileGabriele Bradamante, Zorana Bogojevic, View ORCID ProfileRuben Gutzat, View ORCID ProfileSusanna Streubel, Magdalena Mosiolek, View ORCID ProfileLiam Dolan, View ORCID ProfileOrtrun Mittelsten Scheid
doi: https://doi.org/10.1101/2023.02.09.527713
Mattia Donà
1Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Vienna Biocenter (VBC), 1030 Vienna, Austria
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  • ORCID record for Mattia Donà
  • For correspondence: mattia.dona@gmi.oeaw.ac.at ortrun.mittelsten_scheid@gmi.oeaw.ac.at
Gabriele Bradamante
1Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Vienna Biocenter (VBC), 1030 Vienna, Austria
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  • ORCID record for Gabriele Bradamante
Zorana Bogojevic
1Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Vienna Biocenter (VBC), 1030 Vienna, Austria
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Ruben Gutzat
1Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Vienna Biocenter (VBC), 1030 Vienna, Austria
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  • ORCID record for Ruben Gutzat
Susanna Streubel
1Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Vienna Biocenter (VBC), 1030 Vienna, Austria
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  • ORCID record for Susanna Streubel
Magdalena Mosiolek
1Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Vienna Biocenter (VBC), 1030 Vienna, Austria
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Liam Dolan
1Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Vienna Biocenter (VBC), 1030 Vienna, Austria
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  • ORCID record for Liam Dolan
Ortrun Mittelsten Scheid
1Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Vienna Biocenter (VBC), 1030 Vienna, Austria
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  • ORCID record for Ortrun Mittelsten Scheid
  • For correspondence: mattia.dona@gmi.oeaw.ac.at ortrun.mittelsten_scheid@gmi.oeaw.ac.at
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SUMMARY

Individual cells give rise to diverse cell lineages during the development of multicellular organisms. Understanding the contribution of these lineages to mature organisms is a central question of developmental biology. Several techniques to document cell lineages have been used, from marking single cells with mutations that express a visible marker to generating molecular bar codes by CRISPR-induced mutations and subsequent single-cell analysis. Here, we exploit the mutagenic activity of CRISPR to allow lineage tracing within living plants. Cas9-induced mutations are directed to correct a frameshift mutation that restores expression of a nuclear fluorescent protein, labelling the initial cell and all progenitor cells with a strong signal without modifying other phenotypes of the plants. Spatial and temporal control of Cas9 activity can be achieved using tissue-specific and/or inducible promoters. We provide proof of principle for the function of lineage tracing in two model plants. The conserved features of the components and the versatile cloning system, allowing for easy exchange of promoters, are expected to make the system widely applicable.

SIGNIFICANCE STATEMENT By targeting Cas9 in a tissue- and time-specific way to correct a frameshift mutation, resulting in fluorescence labelling of nuclei, we generated a method for in vivo visual lineage tracing in two model plants. The versatile cloning system makes the system widely applicable in other plants.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted February 10, 2023.
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A versatile CRISPR-based system for lineage tracing in living plants
Mattia Donà, Gabriele Bradamante, Zorana Bogojevic, Ruben Gutzat, Susanna Streubel, Magdalena Mosiolek, Liam Dolan, Ortrun Mittelsten Scheid
bioRxiv 2023.02.09.527713; doi: https://doi.org/10.1101/2023.02.09.527713
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A versatile CRISPR-based system for lineage tracing in living plants
Mattia Donà, Gabriele Bradamante, Zorana Bogojevic, Ruben Gutzat, Susanna Streubel, Magdalena Mosiolek, Liam Dolan, Ortrun Mittelsten Scheid
bioRxiv 2023.02.09.527713; doi: https://doi.org/10.1101/2023.02.09.527713

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