Spatiotemporally controlled genetic perturbation for efficient large-scale studies of cell non-autonomous effects

  1. Andrea Chai
  2. Ana M Mateus
  3. Fazal Oozeer
  4. Rita Sousa-Nunes  Is a corresponding author
  1. King's College London, United Kingdom

Abstract

Studies in genetic model organisms have revealed much about the development and pathology of complex tissues. Most have focused on cell-intrinsic gene functions and mechanisms. Much less is known about how transformed, or otherwise functionally disrupted, cells interact with healthy ones towards a favorable or pathological outcome. This is largely due to technical limitations. We developed new genetic tools in Drosophila melanogaster that permit efficient multiplexed gain- and loss-of-function genetic perturbations with separable spatial and temporal control. Importantly, our novel tool-set is independent of the commonly used GAL4/UAS system, freeing the latter for additional, non-autonomous, genetic manipulations; and is built into a single strain, allowing one-generation interrogation of non-autonomous effects. Altogether, our design opens up efficient genome-wide screens on any deleterious phenotype, once plasmid or genome engineering is used to place the desired miRNA(s) or ORF(s) into our genotype. Specifically, we developed tools to study extrinsic effects on neural tumor growth but the strategy presented has endless applications within and beyond neurobiology, and in other model organisms.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. A source data file has been provided for Figure 2-supplement figure 1 and for Figure 6; FOFO2.0 plasmid sequence has been uploaded; plasmids and transgenic flies will be deposited in stock centres before publication.

Article and author information

Author details

  1. Andrea Chai

    Centre for Developmental Neurobiology, King's College London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  2. Ana M Mateus

    Centre for Developmental Neurobiology, King's College London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  3. Fazal Oozeer

    Centre for Developmental Neurobiology, King's College London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  4. Rita Sousa-Nunes

    Centre for Developmental Neurobiology, King's College London, London, United Kingdom
    For correspondence
    rita.sousa-nunes@kcl.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7401-8081

Funding

Cancer Research UK (Career Development Fellowship)

  • Andrea Chai
  • Ana M Mateus
  • Rita Sousa-Nunes

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Hugo J Bellen, Baylor College of Medicine, United States

Publication history

  1. Received: May 15, 2018
  2. Accepted: November 26, 2018
  3. Accepted Manuscript published: November 27, 2018 (version 1)
  4. Version of Record published: January 4, 2019 (version 2)

Copyright

© 2018, Chai et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Andrea Chai
  2. Ana M Mateus
  3. Fazal Oozeer
  4. Rita Sousa-Nunes
(2018)
Spatiotemporally controlled genetic perturbation for efficient large-scale studies of cell non-autonomous effects
eLife 7:e38393.
https://doi.org/10.7554/eLife.38393

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