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Abstract
Mosaic animals have provided the platform for many fundamental discoveries in developmental biology, cell biology, and other fields. Techniques to produce mosaic animals by mitotic recombination have been extensively developed in Drosophila melanogaster but are less common for other laboratory organisms. Here, we report mosaic analysis by gRNA-induced crossing-over (MAGIC), a new technique for generating mosaic animals based on DNA double-strand breaks produced by CRISPR/Cas9. MAGIC efficiently produces mosaic clones in both somatic tissues and the germline of Drosophila. Further, by developing a MAGIC toolkit for 1 chromosome arm, we demonstrate the method's application in characterizing gene function in neural development and in generating fluorescently marked clones in wild-derived Drosophila strains. Eliminating the need to introduce recombinase-recognition sites into the genome, this simple and versatile system simplifies mosaic analysis in Drosophila and can in principle be applied in any organism that is compatible with CRISPR/Cas9.
Highlights
Mosaic animals contain genetically distinct populations of cells that have arisen from 1 zygote
mosaic analysis by gRNAinduced crossing-over (MAGIC) relies on the action of guide RNA (gRNA)/Cas9 in a proliferating cell during G2 phase to generate a double-strand break (DSB) at a specific position on 1 chromatid of a homologous pair (Fig 1A)
We present here a new technique that we named MAGIC for mosaic analysis based on CRISPR-induced mitotic recombination
Summary
Mosaic animals contain genetically distinct populations of cells that have arisen from 1 zygote. Mosaic animals have historically played important roles in the study of pleiotropic genes, developmental timing, cell lineage, neural wiring, and other complex biological processes. Drosophila has been a major system for generating and studying such mosaics [1], which have led to important discoveries such as developmental compartments [2], cell autonomy [3], and maternal effects of zygotic lethal genes [4]. Mosaic analysis is currently used to study tumor suppressors [5], signaling pathways [6], sleep–wake behaviors [7], cell fates [8], and neuronal lineages [9], among other biological processes.
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