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

Research output: Contribution to journalArticle

Abstract

Genetically encoded effectors are important tools forprobing 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.

Original languageEnglish (US)
Pages (from-to)1410-1421
Number of pages12
JournalCell Reports
Volume10
Issue number8
DOIs
StatePublished - Mar 3 2015

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Drosophila
Exons
Transgenes
Introns
RNA Splice Sites
Animals
Transcription Factors
Genes
Peptides

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Diao, F., Ironfield, H., Luan, H., Diao, F., Shropshire, W. C., Ewer, J., ... White, B. H. (2015). Plug-and-play genetic access to drosophila cell types using exchangeable exon cassettes. Cell Reports, 10(8), 1410-1421. https://doi.org/10.1016/j.celrep.2015.01.059

Plug-and-play genetic access to drosophila cell types using exchangeable exon cassettes. / Diao, Fengqiu; Ironfield, Holly; Luan, Haojiang; Diao, Feici; Shropshire, William C.; Ewer, John; Marr, Elizabeth; Potter, Christopher J; Landgraf, Matthias; White, Benjamin H.

In: Cell Reports, Vol. 10, No. 8, 03.03.2015, p. 1410-1421.

Research output: Contribution to journalArticle

Diao, F, Ironfield, H, Luan, H, Diao, F, Shropshire, WC, Ewer, J, Marr, E, Potter, CJ, Landgraf, M & White, BH 2015, 'Plug-and-play genetic access to drosophila cell types using exchangeable exon cassettes', Cell Reports, vol. 10, no. 8, pp. 1410-1421. https://doi.org/10.1016/j.celrep.2015.01.059
Diao F, Ironfield H, Luan H, Diao F, Shropshire WC, Ewer J et al. Plug-and-play genetic access to drosophila cell types using exchangeable exon cassettes. Cell Reports. 2015 Mar 3;10(8):1410-1421. https://doi.org/10.1016/j.celrep.2015.01.059
Diao, Fengqiu ; Ironfield, Holly ; Luan, Haojiang ; Diao, Feici ; Shropshire, William C. ; Ewer, John ; Marr, Elizabeth ; Potter, Christopher J ; Landgraf, Matthias ; White, Benjamin H. / Plug-and-play genetic access to drosophila cell types using exchangeable exon cassettes. In: Cell Reports. 2015 ; Vol. 10, No. 8. pp. 1410-1421.
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