1. Developmental Biology
  2. Genetics and Genomics

A genome engineering resource to uncover principles of cellular organization and tissue architecture by lipid signalling

  1. Deepti Trivedi
  2. Vinitha CM
  3. Karishma Bisht
  4. Vishnu Janardan
  5. Awadhesh Pandit
  6. Bishal Basak
  7. Shwetha H
  8. Navyashree Ramesh
  9. Padinjat Raghu  Is a corresponding author
  1. National Centre for Biological Sciences, India
https://doi.org/10.7554/eLife.55793
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  1. Deepti Trivedi
  2. Vinitha CM
  3. Karishma Bisht
  4. Vishnu Janardan
  5. Awadhesh Pandit
  6. Bishal Basak
  7. Shwetha H
  8. Navyashree Ramesh
  9. Padinjat Raghu
(2020)
A genome engineering resource to uncover principles of cellular organization and tissue architecture by lipid signalling
eLife 9:e55793.
https://doi.org/10.7554/eLife.55793
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Abstract

Phosphoinositides (PI) are key regulators of cellular organization in eukaryotes and genes that tune PI signalling are implicated in human disease mechanisms. Biochemical analyses and studies in cultured cells have identified a large number of proteins that can mediate PI signalling. However, the role of such proteins in regulating cellular processes in vivo and development in metazoans remains to be understood. Here we describe a set of CRISPR based genome engineering tools that allow the manipulation of each of these proteins with spatial and temporal control during metazoan development. We demonstrate the use of these reagents to deplete a set of 103 proteins individually in the Drosophila eye and identify several new molecules that control eye development. Our work demonstrates the power of this resource in uncovering the molecular basis of tissue homeostasis during normal development and in human disease biology.

Data availability

Full genome sequencing for isogenized Attp40 Stock submitted to NCBI (BioProject ID PRJNA606147). Full genome sequencing for S2R+ cells submitted to NCBI (Bioproject ID PRJNA606149). Images for PI signaling genetic screen saved at Open Source Frame https://osf.io/pt7zu/?view_only=14642fc3a5d74e408fb3766c2555393f

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Article and author information

Author details

  1. Deepti Trivedi

    Cellular Organization and Signalling, National Centre for Biological Sciences, Bangalore, India
    Competing interests
    The authors declare that no competing interests exist.

  2. Vinitha CM

    Cellular Organization and Signalling, National Centre for Biological Sciences, Bangalore, India
    Competing interests
    The authors declare that no competing interests exist.

  3. Karishma Bisht

    Cellular Organization and Signalling, National Centre for Biological Sciences, Bangalore, India
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9088-1141

  4. Vishnu Janardan

    Cellular Organization and Signalling, National Centre for Biological Sciences, Bangalore, India
    Competing interests
    The authors declare that no competing interests exist.

  5. Awadhesh Pandit

    Cellular Organization and Signalling, National Centre for Biological Sciences, Bangalore, India
    Competing interests
    The authors declare that no competing interests exist.

  6. Bishal Basak

    Cellular Organization and Signalling, National Centre for Biological Sciences, Bangalore, India
    Competing interests
    The authors declare that no competing interests exist.

  7. Shwetha H

    Cellular Organization and Signalling, National Centre for Biological Sciences, Bangalore, India
    Competing interests
    The authors declare that no competing interests exist.

  8. Navyashree Ramesh

    Cellular Organization and Signalling, National Centre for Biological Sciences, Bangalore, India
    Competing interests
    The authors declare that no competing interests exist.

  9. Padinjat Raghu

    Cellular Organization and Signalling, National Centre for Biological Sciences, Bangalore, India
    For correspondence
    praghu@ncbs.res.in
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3578-6413

Funding

Department of Biotechnology, Ministry of Science and Technology, India (BT/PRJ3748/GET/l 19/27/2015)

  • Deepti Trivedi
  • Vinitha CM
  • Karishma Bisht
  • Vishnu Janardan
  • Padinjat Raghu

Wellcome-DBT India Alliance (IA/S/14/2/501540)

  • Vinitha CM
  • Karishma Bisht
  • Vishnu Janardan
  • Bishal Basak
  • Padinjat Raghu

National Centre for Biological Sciences (core)

  • Deepti Trivedi
  • Vinitha CM
  • Karishma Bisht
  • Vishnu Janardan
  • Awadhesh Pandit
  • Bishal Basak
  • Padinjat Raghu

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

Reviewing Editor

  1. Jiwon Shim, Hanyang University, Republic of Korea

Publication history

  1. Received: February 6, 2020
  2. Accepted: December 14, 2020
  3. Accepted Manuscript published: December 15, 2020 (version 1)
  4. Version of Record published: December 29, 2020 (version 2)

Copyright

© 2020, Trivedi 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. Deepti Trivedi
  2. Vinitha CM
  3. Karishma Bisht
  4. Vishnu Janardan
  5. Awadhesh Pandit
  6. Bishal Basak
  7. Shwetha H
  8. Navyashree Ramesh
  9. Padinjat Raghu
(2020)
A genome engineering resource to uncover principles of cellular organization and tissue architecture by lipid signalling
eLife 9:e55793.
https://doi.org/10.7554/eLife.55793

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