A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila

Sonal Nagarkar-Jaiswal, Pei Tseng Lee, Megan E. Campbell, Kuchuan Chen, Stephanie Anguiano-Zarate, Manuel Cantu Gutierrez, Theodore Busby, Wen Wen Lin, Yuchun He, Karen L. Schulze, Benjamin W. Booth, Martha Evans-Holm, Koen J T Venken, Robert W. Levis, Allan C. Spradling, Roger A. Hoskins, Hugo J. Bellen

Research output: Contribution to journalArticlepeer-review

Abstract

Here, we document a collection of ∼7434 MiMIC (Minos Mediated Integration Cassette) insertions of which 2854 are inserted in coding introns. They allowed us to create a library of 400 GFP-tagged genes. We show that 72% of internally tagged proteins are functional, and that more than 90% can be imaged in unfixed tissues. Moreover, the tagged mRNAs can be knocked down by RNAi against GFP (iGFPi), and the tagged proteins can be efficiently knocked down by deGradFP technology. The phenotypes associated with RNA and protein knockdown typically correspond to severe loss of function or null mutant phenotypes. Finally, we demonstrate reversible, spatial, and temporal knockdown of tagged proteins in larvae and adult flies. This new strategy and collection of strains allows unprecedented in vivo manipulations in flies for many genes. These strategies will likely extend to vertebrates.

Original languageEnglish (US)
Article numbere05338
JournaleLife
Volume2015
Issue number4
DOIs
StatePublished - Mar 31 2015
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Medicine(all)
  • Neuroscience(all)

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