Abstract
The Notch gene is a key factor in the signaling cascade that allows communication between neighboring cells in many organisms, from worms and insects to humans. The relative simplicity of the Notch pathway in Drosophila, combined with a powerful set of molecular and cytogenetic methods, makes this model attractive for studying the fundamental principles of Notch regulation and functioning. Here, using the CRISPR/Cas9 system in combination with homologous recombination, for the first time at the level of the whole organism, we obtained a directed deletion of the 5′-regulatory region and the first exon of the Notch gene, which were replaced by the attP integration site of the ΦC31 phage. Based on this approach, we obtained and characterized new Notch mutations. Thus, a new powerful tool is provided for studying the genetic regulation of the Notch gene and the organization of chromatin at this locus.
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Acknowledgments
This work was supported by the Russian Science Foundation project #20-14-00074. The authors gratefully acknowledge the resources provided by the “Molecular and Cellular Biology” core facility of the IMCB SB RAS supported by the fundamental scientific research program on the project FWGZ-2021-0014. We thank D.S. Sidorenko at IMCB SB RAS for helpful discussion of this work. We thank the Bloomington Drosophila Stock Center for their kind provision of the fly stocks.
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Andreyenkov, O.V., Volkova, E.I., Andreyenkova, N.G., Demakov, S.A. (2022). Using the CRISPR/Cas9 System for Dissection of Functional Sites of the Notch Gene in Drosophila melanogaster. In: Jia, D. (eds) Notch Signaling Research. Methods in Molecular Biology, vol 2472. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2201-8_1
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DOI: https://doi.org/10.1007/978-1-0716-2201-8_1
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