Cell Reports (Mar 2015)

Plug-and-Play Genetic Access to Drosophila Cell Types using Exchangeable Exon Cassettes

  • Fengqiu Diao,
  • Holly Ironfield,
  • Haojiang Luan,
  • Feici Diao,
  • William C. Shropshire,
  • John Ewer,
  • Elizabeth Marr,
  • Christopher J. Potter,
  • Matthias Landgraf,
  • Benjamin H. White

DOI
https://doi.org/10.1016/j.celrep.2015.01.059
Journal volume & issue
Vol. 10, no. 8
pp. 1410 – 1421

Abstract

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Genetically encoded effectors are important tools for probing cellular function in living animals, but improved methods for directing their expression to specific cell types are required. Here, we introduce a simple, versatile method for achieving cell-type-specific expression of transgenes that leverages the untapped potential of “coding introns” (i.e., introns between coding exons). Our method couples the expression of a transgene to that of a native gene expressed in the cells of interest using intronically inserted “plug-and-play” cassettes (called “Trojan exons”) that carry a splice acceptor site followed by the coding sequences of T2A peptide and an effector transgene. We demonstrate the efficacy of this approach in Drosophila using lines containing suitable MiMIC (Minos-mediated integration cassette) transposons and a palette of Trojan exons capable of expressing a range of commonly used transcription factors. We also introduce an exchangeable, MiMIC-like Trojan exon construct that can be targeted to coding introns using the Crispr/Cas system.