PgmNr Z6093: Establish a Zebrafish genetic mosaic system for single-cell resolution phenotypic analysis of mutant cells.

Authors:
Guoxin Zhang; Laura Fontenas; Sarah Kucenas; Hui Zong


Institutes
University of Virginia, Charlottesville, VA.


Abstract:

Genetic mosaicism is defined by the existence of cell populations with different genotypes in an individual organism. Many human diseases, such as cancer, are caused by genetic mosaicism since cancerous cells harbor genetic mutations that are absent in normal cells within the same person. In research labs, studying mosaic animals reveals cell-autonomous functions of a given gene, particularly suitable for polarity, migration, cell-cell interaction problems. Obviously, the unequivocal labeling of rare mutant cells is the prerequisite for one to analyze them in a genetic mosaic animal. Previously we generated the MADM (mosaic analysis with double markers) system in mice, which generate GFP-labeled mutant cells and RFP-labeled sibling WT cells through site-specific recombinase mediated inter-chromosomal mitotic recombination (Zong 2005 Cell). By comparing green and red cell behaviors in vivo, one can detect even the subtlest phenotypes at the single cell resolution. MADM has been broadly adopted in many fields such as neurobiology (Hippenmeyer 2010 Neuron), developmental biology (Packard 2013 Developmental cell), and cancer biology (Liu 2011 Cell). While we have learned a lot of fascinating biology with the mouse MADM system, Zebrafish model carries great advantages in terms of the transparency of its body and the ease to generate a large, genetically identical population. Based on our experience in establishing the mouse MADM system, we are now in the process of establishing the MADM system in Zebrafish. We envision that the system would not only help deepen our understanding of cellular behaviors in normal development and adult functions, but also enable the establishment of disease models and drug-screening platforms.