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

Elife. 2020 Dec 15;9:e55793. doi: 10.7554/eLife.55793.

Abstract

Phosphoinositides (PI) are key regulators of cellular organization in eukaryotes and genes that tune PI signaling are implicated in human disease mechanisms. Biochemical analyses and studies in cultured cells have identified a large number of proteins that can mediate PI signaling. 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.

Keywords: CRISPR/Cas9; D. melanogaster; Drosophila; developmental biology; eye development; genetics; genome engineering; genomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics*
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Drosophila melanogaster / embryology
  • Drosophila melanogaster / genetics*
  • Eye / embryology*
  • Eye / metabolism
  • Gene Editing / methods
  • Gene Knockout Techniques
  • Genetic Engineering / methods*
  • Genome, Insect / genetics
  • Lipid Metabolism
  • Phosphatidylinositols / metabolism*
  • RNA, Guide / biosynthesis
  • RNA, Guide / genetics
  • Sequence Deletion / genetics
  • Signal Transduction / physiology

Substances

  • Phosphatidylinositols
  • RNA, Guide