Multiplexed drug-based selection and counterselection genetic manipulations in Drosophila

Authors

Nick Matinyan
Mansi S Karkhanis
Yezabel Gonzalez
Antrix Jain
Alexander Saltzman
Anna Malovannaya
Alejandro Sarrion-Perdigones
Herman A Dierick
Koen J T Venken

Publication Date

9-14-2021

Journal

Cell Reports

DOI

10.1016/j.celrep.2021.109700

PMID

34525356

PMCID

PMC8480232

PubMedCentral® Posted Date

9-29-2021

PubMedCentral® Full Text Version

Author MSS

Published Open-Access

no

Keywords

Animals, Animals, Genetically Modified, Chromosomes, Artificial, Bacterial, Drosophila, Drug Resistance, Female, Ganciclovir, Genetic Techniques, Gentamicins, Male, Transgenes

Abstract

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.

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