A genetic toolkit for tagging intronic MiMIC containing genes
- Howard Hughes Medical Institute, Baylor College of Medicine, United States
- Howard Hughes Medical Institute, Carnegie Institution for Science, United States
- Baylor College of Medicine, United States
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.
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© 2015, Nagarkar-Jaiswal et al.
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A genetic toolkit for tagging intronic MiMIC containing genes
eLife 4:e08469.
https://doi.org/10.7554/eLife.08469
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