Tasic, Bosiljka; Miyamichi, Kazunari; Hippenmeyer, SimonISTA ; Dani, Vardhan S.; Zeng, H.; Joo, William; Zong, Hui; Chen-Tsai, Yanru; Luo, Liqun
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.
This work was supported by a National Institutes of Health grant to LL (R01-NS050835). BT was a Damon Runyon Fellow and was supported by the Damon Runyon Cancer Research Foundation Grant DRG-1819-04. KM was supported by the Japan Society for the Promotion of Science program for Research Abroad and Human Frontier Science Program Organization (LT00300/2007-L). SH was supported by postdoctoral fellowships from the European Molecular Biology Organization (ALTF 851-2005), Human Frontier Science Program Organization (LT00805/2006-L), and Swiss National Science Foundation (PA00P3_124160 and PA00P3_136482). HZ is a Pew Scholar in Biomedical Sciences, supported by The Pew Charitable Trusts. LL is an investigator of the Howard Hughes Medical Institute. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Tasic B, Miyamichi K, Hippenmeyer S, et al. Extensions of MADM (Mosaic Analysis with Double Markers) in Mice . PLoS One. 2012;7(3). doi:10.1371/journal.pone.0033332
Tasic, B., Miyamichi, K., Hippenmeyer, S., Dani, V., Zeng, H., Joo, W., … Luo, L. (2012). Extensions of MADM (Mosaic Analysis with Double Markers) in Mice . PLoS One. Public Library of Science. https://doi.org/10.1371/journal.pone.0033332
Tasic, Bosiljka, Kazunari Miyamichi, Simon Hippenmeyer, Vardhan Dani, H. Zeng, William Joo, Hui Zong, Yanru Chen Tsai, and Liqun Luo. “Extensions of MADM (Mosaic Analysis with Double Markers) in Mice .” PLoS One. Public Library of Science, 2012. https://doi.org/10.1371/journal.pone.0033332.
B. Tasic et al., “Extensions of MADM (Mosaic Analysis with Double Markers) in Mice ,” PLoS One, vol. 7, no. 3. Public Library of Science, 2012.
Tasic B, Miyamichi K, Hippenmeyer S, Dani V, Zeng H, Joo W, Zong H, Chen Tsai Y, Luo L. 2012. Extensions of MADM (Mosaic Analysis with Double Markers) in Mice . PLoS One. 7(3).
Tasic, Bosiljka, et al. “Extensions of MADM (Mosaic Analysis with Double Markers) in Mice .” PLoS One, vol. 7, no. 3, Public Library of Science, 2012, doi:10.1371/journal.pone.0033332.