MAZe: a tool for mosaic analysis of gene function in zebrafish.
To trace cell lineages in a developing vertebrate and to observe, in vivo, how behaviors of individual cells are affected by the genes they express, we created a zebrafish line containing a transgene called mosaic analysis in zebrafish (MAZe), built around a self-excising hsp70:Cre cassette. Heat shock triggers Cre recombinase-mediated recombination in a random subset of cells, bringing the transcriptional activator Gal4:VP16 under control of the EF1alpha promoter. Gal4-VP16 then activates expression of a fluorescent protein from an upstream activating sequence (UAS) promoter. Marked clones of cells expressing any desired gene product can be generated by crossing MAZe fish with other lines containing UAS-driven transgenes. The number of clones induced, and their time of origin, could be varied by adjusting heat-shock timing and duration. As an alternative to heat shock, we introduced Cre under a tissue-specific promoter in MAZe fish to generate clones in a designated tissue.
Vertebrate Development Laboratory, Cancer Research UK London Research Institute, London, UK. julian.lewis@cancer.org.uk
Russell T
RT
Collins
Claudia
C
Linker
Julian
J
Lewis
D000818:Animals; D001483:Base Sequence; D002459:Cell Fusion; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D019426:Integrases / Q000502:physiology; D008969:Molecular Sequence Data; D009030:Mosaicism; D032446:Myoblasts / Q000378:metabolism; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D019076:Transgenes; D015027:Zebrafish / Q000235:genetics*
D018840:HSP70 Heat-Shock Proteins; C045073:Cre recombinase; D019426:Integrases