A genetic toolkit for tagging intronic MiMIC containing genes

eLife

Steven Z DeLuca¹  Allan C Spradling¹  Zhongyuan Zuo²  Jiangxing Lv²  Wen-Wen Lin²  Pei-Tseng Lee²  Hongling Pan³  Sonal Nagarkar-Jaiswal³  Hugo J Bellen³ 

[1] Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States;Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States;Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States;

关键词: RMCE;    Drosophila;    GFP protein tagging;    l(2)gl;   

DOI  :  10.7554/eLife.08469

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown