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Citation
Franz, A., Brunner, E., Basler, K. (2017). Generation of genome-modified Drosophila cell lines using SwAP.  Fly 11(4): 303--311.
FlyBase ID
FBrf0237437
Publication Type
Research paper
Abstract

The ease of generating genetically modified animals and cell lines has been markedly increased by the recent development of the versatile CRISPR/Cas9 tool. However, while the isolation of isogenic cell populations is usually straightforward for mammalian cell lines, the generation of clonal Drosophila cell lines has remained a longstanding challenge, hampered by the difficulty of getting Drosophila cells to grow at low densities. Here, we describe a highly efficient workflow to generate clonal Cas9-engineered Drosophila cell lines using a combination of cell pools, limiting dilution in conditioned medium and PCR with allele-specific primers, enabling the efficient selection of a clonal cell line with a suitable mutation profile. We validate the protocol by documenting the isolation, selection and verification of eight independently Cas9-edited armadillo mutant Drosophila cell lines. Our method provides a powerful and simple workflow that improves the utility of Drosophila cells for genetic studies with CRISPR/Cas9.

PubMed ID
PubMed Central ID
PMC5721941 (PMC) (EuropePMC)
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Secondary IDs
    Language of Publication
    English
    Additional Languages of Abstract
    Parent Publication
    Publication Type
    Journal
    Abbreviation
    Fly
    Title
    Fly
    Publication Year
    2007-
    ISBN/ISSN
    1933-6934 1933-6942
    Data From Reference
    Genes (1)
    Cell Lines (1)