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Somatic Clonal Analyses Using FLP/FRT and MARCM System to Understand Notch Signaling Mechanism and Its Regulation

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Notch Signaling Research

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2472))

Abstract

Notch signaling regulates an array of developmental decisions and has been implicated in a multitude of diseases, including cancer over the past a few decades. The simplicity and versatility of the Notch pathway in Drosophila make it an ardent system to study Notch biology, its regulation, and functions. In this chapter, we highlight the use of two powerful techniques, namely, FLP/FRT and MARCM in the study of Notch signaling. These mosaic analysis techniques are powerful tools to analyze gene functions in different biological processes. The section briefly explains the principle and the protocols with suitable examples.

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References

  1. Andersson ER, Sandberg R, Lendahl U (2011) Notch signaling: simplicity in design, versatility in function. Development 138:3593–3612

    Article  CAS  Google Scholar 

  2. Artavanis-Tsakonas S, Rand MD, Lake RJ (1999) Notch signaling: cell fate control and signal integration in development. Science 284:770–776

    Article  CAS  Google Scholar 

  3. Fortini ME (2009) Notch signaling: the core pathway and its posttranslational regulation. Dev Cell 16:633–647

    Article  CAS  Google Scholar 

  4. Kopan R, Ilagan MXG (2009) The canonical Notch signaling pathway: unfolding the activation mechanism. Cell 137:216–233

    Article  CAS  Google Scholar 

  5. Siebel C, Lendahl U (2017) Notch signaling in development, tissue homeostasis, and disease. Physiol Rev 97:1235–1294

    Article  CAS  Google Scholar 

  6. Zacharioudaki E, Bray SJ (2014) Tools and methods for studying Notch signaling in Drosophila melanogaster. Methods 68:173–182

    Article  CAS  Google Scholar 

  7. Golic KG, Lindquist S (1989) The FLP recombinase of yeast catalyzes site-specific recombination in the Drosophila genome. Cell 59:499–509

    Article  CAS  Google Scholar 

  8. Xu T, Rubin GM (1993) Analysis of genetic mosaics in developing and adult Drosophila tissues. Development 117:1223–1237

    Article  CAS  Google Scholar 

  9. Brand AH, Perrimon N (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118:401–415

    Article  CAS  Google Scholar 

  10. Lee T, Luo L (1999) Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis. Neuron 22:451–461

    Article  CAS  Google Scholar 

  11. Lee T, Luo L (2001) Mosaic analysis with a repressible cell marker (MARCM) for Drosophila neural development. Trends Neurosci 24:251–254

    Article  CAS  Google Scholar 

  12. Sachan N, Mishra AK, Mutsuddi M et al (2015) Chip physically interacts with Notch and their stoichiometry is critical for Notch function in wing development and cell proliferation in Drosophila. Biochim Biophys Acta Gen Subj 1850:802–812

    Article  CAS  Google Scholar 

  13. Sachan N, Mishra AK, Mutsuddi M et al (2013) The Drosophila importin-α3 is required for nuclear import of Notch in vivo and it displays synergistic effects with Notch receptor on cell proliferation. PLoS One 8:e68247

    Article  CAS  Google Scholar 

  14. Mukherjee A, Veraksa A, Bauer A et al (2005) Regulation of Notch signalling by non-visual β-arrestin. Nat Cell Biol 7:1191–1201

    Article  CAS  Google Scholar 

  15. Mishra AK, Sachan N, Mutsuddi M et al (2014) TRAF6 is a novel regulator of Notch signaling in Drosophila melanogaster. Cell Signal 26:3016–3026

    Article  CAS  Google Scholar 

  16. Paul MS, Dutta D, Singh A et al (2018) Regulation of Notch signaling in the developing Drosophila eye by a T-box containing transcription factor, Dorsocross. Genesis 56(10):e23251

    Article  CAS  Google Scholar 

  17. Singh A, Dutta D, Paul MS et al (2018) Pleiotropic functions of the chromodomain-containing protein hat-trick during oogenesis in Drosophila melanogaster. G3 (Bethesda) 8:1067–1077

    Article  CAS  Google Scholar 

  18. Singh A, Paul MS, Dutta D et al (2019) Regulation of Notch signaling by the chromatin-modeling protein Hat-trick. Development 146:dev170837

    Article  CAS  Google Scholar 

  19. Sharma V, Mutsuddi M, Mukherjee A (2021) Deltex cooperates with TRAF6 to promote apoptosis and cell migration through Eiger-independent JNK activation in Drosophila. Cell Biol Int 45:686–700

    Article  CAS  Google Scholar 

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Acknowledgments

We extend our sincere thanks to the Bloomington Stock Centre and Developmental studies Hybridoma Bank (DSHB) for fly stocks and some of the antibodies used in our studies. We also acknowledge the confocal facility of DBT-BHU-ISLS, Banaras Hindu University for microscopy assistance. This work was supported by grants from Department of Science and Technology (DST), Government of India and Institute of Eminence Scheme, Banaras Hindu University, India. Fellowship support to Vartika Sharma was provided by the Council of Scientific and Industrial Research (CSIR), Government of India. Artwork is created with BioRender (BioRender.com).

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Authors and Affiliations

  1. Department of Molecular and Human Genetics, Institute of Science, Banaras Hindu University, Varanasi, Uttar Pradesh, India

    Vartika Sharma, Mousumi Mutsuddi & Ashim Mukherjee

  2. Department of Cell Biology, NYU Grossman School of Medicine, New York, NY, USA

    Nalani Sachan

Authors
  1. Vartika Sharma
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  2. Nalani Sachan
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  3. Mousumi Mutsuddi
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  4. Ashim Mukherjee
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Corresponding author

Correspondence to Ashim Mukherjee .

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Editors and Affiliations

  1. Department of Biology, Georgia Southern University, Statesboro, GA, USA

    Dongyu Jia

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Sharma, V., Sachan, N., Mutsuddi, M., Mukherjee, A. (2022). Somatic Clonal Analyses Using FLP/FRT and MARCM System to Understand Notch Signaling Mechanism and Its Regulation. 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_8

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  • DOI: https://doi.org/10.1007/978-1-0716-2201-8_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2200-1

  • Online ISBN: 978-1-0716-2201-8

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