Plug-and-play Genetic Access to Drosophila Cell Types Using Exchangeable Exon Cassettes

Cell Rep. 2015 Mar 3;10(8):1410-21. doi: 10.1016/j.celrep.2015.01.059. Epub 2015 Feb 26.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5' Untranslated Regions
  • Animals
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics
  • Drosophila / genetics
  • Drosophila / metabolism*
  • Drosophila Proteins / genetics*
  • Drosophila Proteins / metabolism
  • Exons
  • Introns
  • RNA Splice Sites
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Transgenes / genetics
  • Transgenes / physiology

Substances

  • 5' Untranslated Regions
  • Drosophila Proteins
  • GAL4 protein, Drosophila
  • RNA Splice Sites
  • Transcription Factors