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A protocol for mosaic analysis with a repressible cell marker (MARCM) in Drosophila

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Abstract

Mosaic analysis with a repressible cell marker (MARCM) is a genetic technique used in Drosophila to label single cells or multiple cells sharing a single progenitor. Labeled homozygous mutant cells can be generated in an otherwise unlabeled heterozygous animal. Mutant or wild-type labeled cells can also be made to express one or more transgenes. Major applications of MARCM include (i) lineage analysis, (ii) investigating gene function in single or small populations of cells and (iii) neuronal circuit tracing. Our laboratory uses MARCM primarily to label and genetically manipulate neurons; however, this protocol can be adapted to any cell of interest. The protocol involves generating two fly stocks with the necessary genetic elements for MARCM analysis and subsequently generating MARCM clones. Labeled clones can be followed in live and fixed tissues for high-resolution analysis of wild-type or genetically manipulated cells.

NOTE: In the PDF version of this article initially published online, the first “FRT” and the “Mutation” labels in Figure 1b were transposed. In both the PDF and HTML versions, “mutant” was omitted from the label on the right, which should read “Labeled homozygous mutant daughter cell”. The figure has been corrected in all versions of the article.

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Figure 1: Schematic representation of the GAL4–UAS system with GAL80 and the MARCM genetic system.
Figure 2: Cross between a MARCM-ready and a FRT, mutant fly for MARCM analysis.
Figure 3: MARCM example in which the birth order of projection neurons predicts their dendritic-projection pattern.
Figure 4: MARCM example with high-resolution axon phenotypes in single-cell mutant and single-cell rescue experiments.

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  • 15 February 2007

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Acknowledgements

We thank members of our laboratory for their helpful comments on this protocol. Research in our laboratory has been generously supported by grants from the National Institutes of Health and, more recently, from the Howard Hughes Medical Institute, of which L.L. is an investigator.

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Correspondence to Liqun Luo.

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Wu, J., Luo, L. A protocol for mosaic analysis with a repressible cell marker (MARCM) in Drosophila. Nat Protoc 1, 2583–2589 (2006). https://doi.org/10.1038/nprot.2006.320

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