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A genetic toolkit for tagging intronic MiMIC containing genes

  1. Sonal Nagarkar-Jaiswal
  2. Steven Z DeLuca
  3. Pei-Tseng Lee
  4. Wen-Wen Lin
  5. Hongling Pan
  6. Zhongyuan Zuo
  7. Jiangxing Lv
  8. Allan C Spradling
  9. Hugo J Bellen  Is a corresponding author
  1. Howard Hughes Medical Institute, Baylor College of Medicine, United States
  2. Howard Hughes Medical Institute, Carnegie Institution for Science, United States
  3. Baylor College of Medicine, United States
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Cite this article as: eLife 2015;4:e08469 doi: 10.7554/eLife.08469

Abstract

Previously we described a large collection of MiMICs that contain two phiC31 recombinase target sites and allow the generation of a new exon that encodes a protein tag when the MiMIC (Minos Mediated Integration Cassette) is inserted in a codon intron (Nagarkar-Jaiswal et al., 2015). These modified genes permit numerous applications including assessment of protein expression pattern, identification of protein interaction partners by immunoprecipitation followed by mass spec, and reversible removal of the tagged protein in any tissue. At present, these conversions remain time and labor-intensive as they require embryos to be injected with plasmid DNA containing the exon tag. Here we describe a simple and reliable genetic strategy to tag genes/proteins that contain MiMIC insertions using an integrated exon encoding GFP flanked by FRT sequences. We document the efficiency and tag 60 mostly uncharacterized genes.

Article and author information

Author details

  1. Sonal Nagarkar-Jaiswal

    Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Steven Z DeLuca

    Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Pei-Tseng Lee

    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Wen-Wen Lin

    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Hongling Pan

    Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Zhongyuan Zuo

    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Jiangxing Lv

    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Allan C Spradling

    Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Hugo J Bellen

    Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States
    For correspondence
    hbellen@bcm.edu
    Competing interests
    The authors declare that no competing interests exist.

Reviewing Editor

  1. Mani Ramaswami, Trinity College Dublin, Ireland

Publication history

  1. Received: May 5, 2015
  2. Accepted: June 22, 2015
  3. Accepted Manuscript published: June 23, 2015 (version 1)
  4. Version of Record published: July 13, 2015 (version 2)

Copyright

© 2015, Nagarkar-Jaiswal et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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