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MARCM
Mosaic analysis with a repressible cell marker, or MARCM, is a genetics technique for creating individually labeled homozygous cells in an otherwise heterozygous Drosophila melanogaster. It has been a crucial tool in studying the development of the drosophila nervous system. This technique relies on recombination during mitosis mediated by FLP-FRT Recombination. As one copy of a gene, provided by the balancer chromosome, is often enough to rescue a mutant phenotype, MARCM clones can be used to study a mutant phenotype in an otherwise wildtype animal.
== MARCM Crosses ==
In order to label small populations of cells from a common progenitor, MARCM uses the GAL4-UAS system. A marker gene such as GFP is placed under control of a UAS promoter. GAL4 is ubiquitously expressed in these flies, thus driving marker expression. In addition, GAL80 is driven by a strong promoter such as tubP. GAL80 is an inhibitor of GAL4, and will suppress GFP expression under normal conditions. This tubP-GAL80 element is placed distal to an FRT site. A second FRT site is placed in trans to the GAL80 site, usually with a gene or mutation of interest distal to it. Finally, FLP recombinase is driven by an inducible promoter such as heat shock.
When FLP transcription is induced, it will recombine the chromosomes at the 2 FRT sites in cells undergoing mitosis. These cells will divide into 2 homozygous daughter cells- one carrying both GAL80 elements, and one carrying none. The daughter cell lacking GAL80 will be labeled due to expression of the marker via the GAL4-UAS system. All subsequent daughter cells from this progenitor will also express the marker.
Labs will often have MARCM-ready lines which have the inducible FLP, GAL80 distal to a FRT site, GAL4, and UAS-Marker. These can be readily crossed with flies that have a mutation of interest distal to a FRT site.
抄文引用元・出典: フリー百科事典『 ウィキペディア（Wikipedia）』
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