Wu Joy S, Luo Liqun
Howard Hughes Medical Institute, Department of Biological Sciences, Neurosciences Program, Stanford University, Stanford, California 94305-5020, USA.
Nat Protoc. 2006;1(6):2583-9. doi: 10.1038/nprot.2006.320.
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.
利用可抑制细胞标记进行的镶嵌分析（MARCM）是一种在果蝇中使用的遗传技术，用于标记单个细胞或共享单个祖细胞的多个细胞。可以在其他未标记的杂合动物中产生标记的纯合突变细胞。突变或野生型标记细胞也可以被诱导表达一个或多个转基因。MARCM的主要应用包括：（i）谱系分析；（ii）研究单个或小细胞群体中的基因功能；（iii）神经回路追踪。我们实验室主要使用MARCM来标记和遗传操纵神经元；然而，该方案可适用于任何感兴趣的细胞。该方案包括产生两种具有MARCM分析所需遗传元件的果蝇品系，随后产生MARCM克隆。标记的克隆可以在活体和固定组织中进行追踪，以便对野生型或遗传操纵的细胞进行高分辨率分析。注意：在本文最初在线发表的PDF版本中，图1b中的第一个“FRT”和“突变”标签位置颠倒。在PDF和HTML版本中，右侧标签中都遗漏了“突变体”，应改为“标记的纯合突变子代细胞”。本文所有版本中的图都已更正。
