One-step CRISPR-Cas9 protocol for the generation of plug & play conditional knockouts in Drosophila melanogaster

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Abstract

This one-step method generates, for any locus, a conditional knockout allele in . The allele carries a bright fluorescent marker for easy screening and an landing site for subsequent genetic manipulations. After removing the selectable marker with Cre, the site can be used to insert DNA fragments expressing tagged or mutant alleles to determine protein localization and function in a tissue- and stage-specific manner. Only a single round of CRISPR-Cas9-mediated editing is required. For complete details on the use and execution of this protocol, please refer to the example in Yu et al. (2020).