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

Extensions of MADM (mosaic analysis with double markers) in mice.
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DOI:
10.1371/journal.pone.0033332
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发表时间:
2012
期刊:
影响因子:
3.7
通讯作者:
Luo L
Luo L
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Tasic B;Miyamichi K;Hippenmeyer S;Dani VS;Zeng H;Joo W;Zong H;Chen-Tsai Y;Luo L

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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.
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