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A genetic toolkit for tagging intronic MiMIC containing genes

Authors: Allan C. Spradling; Steven Z DeLuca; Hugo J. Bellen; Zhongyuan Zuo; Sonal Nagarkar-Jaiswal; Jiangxing Lv; Hongling Pan; +2 Authors

A genetic toolkit for tagging intronic MiMIC containing genes

Abstract

Previously, we described a large collection of Minos-Mediated Integration Cassettes (MiMICs) that contain two phiC31 recombinase target sites and allow the generation of a new exon that encodes a protein tag when the MiMIC is inserted in a codon intron (<xref ref-type="bibr" rid="bib17">Nagarkar-Jaiswal et al., 2015</xref>). 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. In this study, 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.

Keywords

QH301-705.5, Science, Genetic Vectors, Green Fluorescent Proteins, Transposases, l(2)gl, Genes, Reporter, Animals, Biology (General), RMCE, Recombination, Genetic, GFP protein tagging, Staining and Labeling, Q, R, Cell Biology, Artificial Gene Fusion, Mutagenesis, Insertional, Gene Targeting, Medicine, Drosophila, Plasmids

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    This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
131
Top 1%
Top 10%
Top 1%
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gold