title: Extensions of MADM (mosaic analysis with double markers) in mice.
abstract: 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.
authors.affiliation: Department of Biology, Howard Hughes Medical Insitute, Stanford University, California, United States of America. bosiljka.tasic@gmail.com
authors.forename: Bosiljka
authors.initials: B
authors.lastname: Tasic
authors.forename: Kazunari
authors.initials: K
authors.lastname: Miyamichi
authors.forename: Simon
authors.initials: S
authors.lastname: Hippenmeyer
authors.forename: Vardhan S
authors.initials: VS
authors.lastname: Dani
authors.forename: Hong
authors.initials: H
authors.lastname: Zeng
authors.forename: William
authors.initials: W
authors.lastname: Joo
authors.forename: Hui
authors.initials: H
authors.lastname: Zong
authors.forename: Yanru
authors.initials: Y
authors.lastname: Chen-Tsai
authors.forename: Liqun
authors.initials: L
authors.lastname: Luo
mesh_terms: D000782:Aneuploidy; D000818:Animals; D002503:Centromere / Q000235:genetics; D033481:Chromosomes, Mammalian / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008957:Models, Genetic; D009154:Mutation; D011995:Recombination, Genetic; D019076:Transgenes / Q000235:genetics*; D014178:Translocation, Genetic; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase