Abstract
Cells differentiate from undifferentiated precursors in order to establish the tissues of vascular plants. The different cell types and stem cells are first specified in the early embryo. How cell type specification is instructed by transcriptional control on a genome-wide level is poorly understood. A major hurdle has been the technical challenge associated with obtaining cellular transcriptomes in this inaccessible tissue. Recently, we adapted a two-component genetic labeling system called INTACT to isolate nuclei and generate a microarray-based expression atlas of the cell types in the early Arabidopsis thaliana embryo. Here we present a step-by-step description of the adapted INTACT protocol, as well as the approach to generate transcriptomic profiles. This protocol has been adapted to account for using seeds with embryos of various developmental stages as a starting material, and the relatively few cell type-specific nuclei that can be isolated from embryos.
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References
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Acknowledgments
The authors thank Tatyana Radoeva and Thomas Nakel for providing the Arabidopsis seed image in Fig. 1a and for image editing in Fig. 1b, respectively. This work was supported by the Federation of European Biochemical Societies (FEBS) to J.P. and by the European Research Council (ERC; Starting Grant “CELLPATTERN”; Contract number 281573) and ERA-CAPS (EURO-PEC; 849.13.006) to D.W.
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Palovaara, J., Weijers, D. (2020). Cell Type-Specific Transcriptomics in the Plant Embryo Using an Adapted INTACT Protocol. In: Bayer, M. (eds) Plant Embryogenesis. Methods in Molecular Biology, vol 2122. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0342-0_11
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DOI: https://doi.org/10.1007/978-1-0716-0342-0_11
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