A genetic toolkit for tagging intronic MiMIC containing genes
Sonal Nagarkar-Jaiswal,
Steven Z DeLuca,
Pei-Tseng Lee,
Wen-Wen Lin,
Hongling Pan,
Zhongyuan Zuo,
Jiangxing Lv,
Allan C Spradling,
Hugo J Bellen
Affiliations
Sonal Nagarkar-Jaiswal
Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States
Steven Z DeLuca
Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States
Pei-Tseng Lee
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
Wen-Wen Lin
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
Hongling Pan
Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States
Zhongyuan Zuo
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
Jiangxing Lv
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
Allan C Spradling
Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States
Hugo J Bellen
Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States; Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States; Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital, Houston, United States; Department of Neuroscience, Baylor College of Medicine, Houston, United States; Program in Developmental Biology, Baylor College of Medicine, Houston, United States
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.
