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Extensions of MADM (mosaic analysis with double markers) in mice.

PloS one | 2012

Mosaic Analysis with Double Markers (MADM) is a method for generating genetically mosaic mice, in which sibling mutant and wild-type cells are labeled with different fluorescent markers. It is a powerful tool that enables analysis of gene function at the single cell level in vivo. It requires transgenic cassettes to be located between the centromere and the mutation in the gene of interest on the same chromosome. Here we compare procedures for introduction of MADM cassettes into new loci in the mouse genome, and describe new approaches for expanding the utility of MADM. We show that: 1) Targeted homologous recombination outperforms random transgenesis in generation of reliably expressed MADM cassettes, 2) MADM cassettes in new genomic loci need to be validated for biallelic and ubiquitous expression, 3) Recombination between MADM cassettes on different chromosomes can be used to study reciprocal chromosomal deletions/duplications, and 4) MADM can be modified to permit transgene expression by combining it with a binary expression system. The advances described in this study expand current, and enable new and more versatile applications of MADM.

Pubmed ID: 22479386 RIS Download

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Associated grants

  • Agency: NINDS NIH HHS, United States
    Id: R01 NS050835
  • Agency: Howard Hughes Medical Institute, United States
  • Agency: NINDS NIH HHS, United States
    Id: R01-NS050835

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RRID:SCR_004286

Commercial antibody vendor which supplies antibodies and other products to life science researchers.

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