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

TLDR

These strategies may extend to vertebrates. We cataloged ~7,434 MiMIC insertions, including 2,854 in coding introns. The library of ~400 GFP‑tagged genes, with 72 % functional and >90 % imaged, permits RNAi and deGradFP‑mediated knockdown of tagged mRNAs and proteins, producing loss‑of‑function phenotypes and enabling reversible, spatial, and temporal protein depletion in larvae and adults, thereby providing unprecedented in‑vivo manipulation of many genes.

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.