Multiplexed drug-based selection and counterselection genetic manipulations in Drosophila
Nick Matinyan,
Mansi S. Karkhanis,
Yezabel Gonzalez,
Antrix Jain,
Alexander Saltzman,
Anna Malovannaya,
Alejandro Sarrion-Perdigones,
Herman A. Dierick,
Koen J.T. Venken
Affiliations
Nick Matinyan
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA; Integrative Molecular Biomedical Sciences Graduate Program, Baylor College of Medicine, Houston, TX 77030, USA
Mansi S. Karkhanis
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, USA
Yezabel Gonzalez
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA
Antrix Jain
Advanced Technology Cores, Mass Spectrometry Proteomics, Baylor College of Medicine, Houston, TX 77030, USA
Alexander Saltzman
Advanced Technology Cores, Mass Spectrometry Proteomics, Baylor College of Medicine, Houston, TX 77030, USA
Anna Malovannaya
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA; Advanced Technology Cores, Mass Spectrometry Proteomics, Baylor College of Medicine, Houston, TX 77030, USA; Dan L. Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX 77030, USA
Alejandro Sarrion-Perdigones
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA
Herman A. Dierick
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, USA; Department of Neuroscience, Baylor College of Medicine, Houston, TX 77030, USA; Corresponding author
Koen J.T. Venken
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA; Integrative Molecular Biomedical Sciences Graduate Program, Baylor College of Medicine, Houston, TX 77030, USA; Dan L. Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX 77030, USA; Department of Pharmacology and Chemical Biology, Baylor College of Medicine, Houston, TX 77030, USA; McNair Medical Institute at The Robert and Janice McNair Foundation, Baylor College of Medicine, Houston, TX 77030, USA; Corresponding author
Summary: The power of Drosophila melanogaster as a model system relies on tractable germline genetic manipulations. Despite Drosophila’s expansive genetics toolbox, such manipulations are still accomplished one change at a time and depend predominantly on phenotypic screening. We describe a drug-based genetic platform consisting of four selection and two counterselection markers, eliminating the need to screen for modified progeny. These markers work reliably individually or in combination to produce specific genetic outcomes. We demonstrate three example applications of multiplexed drug-based genetics by generating (1) transgenic animals, expressing both components of binary overexpression systems in a single transgenesis step; (2) dual selectable and counterselectable balancer chromosomes; and (3) selectable, fluorescently tagged P[acman] bacterial artificial chromosome (BAC) strains. We perform immunoprecipitation followed by proteomic analysis on one tagged BAC line, demonstrating our platform’s applicability to biological discovery. Lastly, we provide a plasmid library resource to facilitate custom transgene design and technology transfer to other model systems.
