A genome engineering resource to uncover principles of cellular organization and tissue architecture by lipid signalling
- National Centre for Biological Sciences, India
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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Full genome sequencing for isogenized Attp40 StockNCBI, BioProject ID PRJNA606147.
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Full genome sequencing for S2R+ cellsNCBI, BioProject ID PRJNA606149.
Article and author information
Author details
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
- Jiwon Shim, Hanyang University, Republic of Korea
Version history
- Received: February 6, 2020
- Accepted: December 14, 2020
- Accepted Manuscript published: December 15, 2020 (version 1)
- 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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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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