title: 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 (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. 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.
authors.affiliation: Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States.
authors.forename: Sonal
authors.initials: S
authors.lastname: Nagarkar-Jaiswal
authors.affiliation: Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States.
authors.forename: Steven Z
authors.initials: SZ
authors.lastname: DeLuca
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States.
authors.forename: Pei-Tseng
authors.initials: PT
authors.lastname: Lee
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States.
authors.forename: Wen-Wen
authors.initials: WW
authors.lastname: Lin
authors.affiliation: Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States.
authors.forename: Hongling
authors.initials: H
authors.lastname: Pan
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States.
authors.forename: Zhongyuan
authors.initials: Z
authors.lastname: Zuo
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States.
authors.forename: Jiangxing
authors.initials: J
authors.lastname: Lv
authors.affiliation: Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States.
authors.forename: Allan C
authors.initials: AC
authors.lastname: Spradling
authors.affiliation: Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States.
authors.forename: Hugo J
authors.initials: HJ
authors.lastname: Bellen
mesh_terms: D000818:Animals; D019677:Artificial Gene Fusion; D004330:Drosophila; D018390:Gene Targeting / Q000379:methods*; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D016254:Mutagenesis, Insertional; D010957:Plasmids; D011995:Recombination, Genetic; D013194:Staining and Labeling / Q000379:methods*; D019895:Transposases / Q000378:metabolism
chemical_list: D049452:Green Fluorescent Proteins; C473499:Minos transposase; D019895:Transposases
keywords: D. melanogaster; Drosophila; GFP protein tagging; RMCE; cell biology; l(2)gl; neuroscience