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

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 markin...

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Bibliographic Details
Published in:The Plant journal : for cell and molecular biology 2023-09, Vol.115 (5), p.1169-1184
Main Authors: Donà, Mattia, Bradamante, Gabriele, Bogojevic, Zorana, Gutzat, Ruben, Streubel, Susanna, Mosiolek, Magdalena, Dolan, Liam, Mittelsten Scheid, Ortrun
Format: Article
Language:English
Subjects:
Arabidopsis thaliana
Cells (biology)
Cloning
CRISPR
Developmental biology
Fluorescence
fluorescence labelling
frameshift correction
Frameshift mutation
in vivo
Labeling
lineage tracing
Marchantia polymorpha
Mutation
Phenotypes
Progenitor cells
Promoters
Tracing
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Summary: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 with a single reporter. 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.
ISSN:0960-7412
1365-313X
DOI:10.1111/tpj.16378