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

Plug-and-play genetic access to drosophila cell types using exchangeable exon cassettes.

Author information

1
Laboratory of Molecular Biology, National Institute of Mental Health, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD 20892, USA.
2
Department of Zoology, University of Cambridge, Downing Street, Cambridge CB2 3EJ, UK.
3
Centro Interdisciplinario de Neurociencia, Universidad de Valparaiso, Pasaje Harrington 287, Playa Ancha, Valparaiso, Chile.
4
The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, 855 North Wolfe Street, Baltimore, MD 21205, USA.
5
Laboratory of Molecular Biology, National Institute of Mental Health, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD 20892, USA. Electronic address: benjaminwhite@mail.nih.gov.

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.

PMID:
25732830
PMCID:
PMC4373654
DOI:
10.1016/j.celrep.2015.01.059
[Indexed for MEDLINE]
Free PMC Article

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