Mosaic Analysis with Double Markers (MADM) offers a unique approach to visualize and concomitantly manipulate genetically-defined cells in mice with single-cell resolution. MADM applications include the analysis of lineage; single-cell morphology and physiology; genomic imprinting phenotypes; and dissection of cell-autonomous gene functions in vivo in health and disease. Yet, MADM could only be applied to <25% of all mouse genes on select chromosomes thus far. To overcome this limitation, we generated transgenic mice with knocked-in MADM cassettes near the centromeres of all 19 autosomes and validated their use across organs. With this resource, >96% of the entire mouse genome can now be subjected to single-cell genetic mosaic analysis. Beyond proof-of-principle, we applied our MADM library to systematically trace sister chromatid segregation in distinct mitotic cell lineages. We found striking chromosome-specific biases in segregation patterns, reflecting a putative mechanism for the asymmetric segregation of genetic determinants in somatic stem cell division.
We thank the Bioimaging-, Life Science- and Pre-Clinical Facilities at IST Austria; MP. Postiglione, C. Simbriger, K. Valskova, C. Schwayer, T. Hussain, M. Pieber, and Victoria Wimmer for initial experiments, technical support and/or assistance; M. Sixt, and all members of the Hippenmeyer lab for discussion; and W. Zhong for sharing Nestin-Cre mice. This work was supported by National Institutes of Health grants (R01-NS050580 to L.L. and F32MH096361 to L.A.S.). L.L. is an investigator of HHMI. N.A. received support from FWF Firnberg-Programm (T 1031). A.H. is a recipient of a DOC Fellowship (24812) of the Austrian Academy of Sciences. This work also received support from IST Austria institutional funds; FWF SFB F78 to S.H.; the People Programme (Marie Curie Actions) of the European Union’s Seventh Framework Programme (FP7/2007-2013) under REA grant agreement No 618444 to S.H., and the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No 725780 LinPro) to S.H.
Contreras X, Davaatseren A, Amberg N, et al. A genome-wide library of MADM mice for single-cell genetic mosaic analysis. bioRxiv. 2020. doi:10.1101/2020.06.05.136192
Contreras, X., Davaatseren, A., Amberg, N., Hansen, A. H., Sonntag, J., Andersen, L., … Hippenmeyer, S. (2020). A genome-wide library of MADM mice for single-cell genetic mosaic analysis. bioRxiv. Cold Spring Harbor Laboratory. https://doi.org/10.1101/2020.06.05.136192
Contreras, Ximena, Amarbayasgalan Davaatseren, Nicole Amberg, Andi H Hansen, Johanna Sonntag, Lill Andersen, Tina Bernthaler, et al. “A Genome-Wide Library of MADM Mice for Single-Cell Genetic Mosaic Analysis.” BioRxiv. Cold Spring Harbor Laboratory, 2020. https://doi.org/10.1101/2020.06.05.136192.
X. Contreras et al., “A genome-wide library of MADM mice for single-cell genetic mosaic analysis,” bioRxiv. Cold Spring Harbor Laboratory, 2020.
Contreras X, Davaatseren A, Amberg N, Hansen AH, Sonntag J, Andersen L, Bernthaler T, Heger A-M, Johnson R, Schwarz LA, Luo L, Rülicke T, Hippenmeyer S. 2020. A genome-wide library of MADM mice for single-cell genetic mosaic analysis. bioRxiv, 10.1101/2020.06.05.136192.
Contreras, Ximena, et al. “A Genome-Wide Library of MADM Mice for Single-Cell Genetic Mosaic Analysis.” BioRxiv, Cold Spring Harbor Laboratory, 2020, doi:10.1101/2020.06.05.136192.