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Vector and parameters for targeted transgenic RNA interference in Drosophila melanogaster

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Abstract

The conditional expression of hairpin constructs in Drosophila melanogaster has emerged in recent years as a method of choice in functional genomic studies. To date, upstream activating site–driven RNA interference constructs have been inserted into the genome randomly using P-element–mediated transformation, which can result in false negatives due to variable expression. To avoid this problem, we have developed a transgenic RNA interference vector based on the phiC31 site-specific integration method.

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Figure 1: Valium is an effective vector for transgenic RNAi.
Figure 2: Parameters of in vivo RNAi using Valium.

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Acknowledgements

We thank G. Rubin, C. Zuker, K. Moses and B. Mathey-Prevot for critical input on the project, B. Dickson (IMP, Vienna) for the gift of UAS-Dcr2 and F. Karch (University of Geneva) for nanos-integrase. M.M. is a fellow of the Jane Coffin Childs Memorial Fund. M.B. is supported by R01 GM067761 from the US National Institute of General Medical Sciences. This work was supported in part by the Janelia Farm Visitor program. N.P. is an investigator of the Howard Hughes Medical Institute.

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Author notes

  1. Jian-Quan Ni, Michele Markstein and Richard Binari: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, 02175, Massachusetts, USA

    Jian-Quan Ni, Michele Markstein, Richard Binari, Lu-Ping Liu, Christians Villalta, Matthew Booker & Norbert Perrimon

  2. Howard Hughes Medical Institute, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, 02175, Massachusetts, USA

    Jian-Quan Ni, Richard Binari, Lu-Ping Liu, Christians Villalta & Norbert Perrimon

  3. Howard Hughes Medical Institute, Janelia Farm Research Campus, 19700 Helix Drive, Ashburn, 20147, Virginia, USA

    Barret Pfeiffer

  4. Pediatric Surgical Research Labs, Massachusetts General Hospital, Harvard Medical School, 185 Cambridge Street, CPZN6.222, Boston, 02114, Massachusetts, USA

    Lizabeth Perkins

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  1. Jian-Quan Ni
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  2. Michele Markstein
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  5. Lu-Ping Liu
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  6. Christians Villalta
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  8. Lizabeth Perkins
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  9. Norbert Perrimon
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Corresponding author

Correspondence to Norbert Perrimon.

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Supplementary Table 1, Supplementary Methods, Supplementary Figure 1 and Supplementary Note (PDF 1804 kb)

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Ni, JQ., Markstein, M., Binari, R. et al. Vector and parameters for targeted transgenic RNA interference in Drosophila melanogaster. Nat Methods 5, 49–51 (2008). https://doi.org/10.1038/nmeth1146

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  • DOI: https://doi.org/10.1038/nmeth1146

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