MAZe: a new tool for mosaic analysis of gene function in zebrafish

R Collins; C Linker; J Lewis

doi:10.1038/nmeth.1423

MAZe: a new tool for mosaic analysis of gene function in zebrafish

R Collins, C Linker, J Lewis

Research output: Contribution to journal › Article › peer-review

55 Citations (Scopus)

Abstract

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 EF1α 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.

Original language	English
Pages (from-to)	219 - 223
Number of pages	5
Journal	NATURE METHODS
Volume	7
Issue number	3
DOIs	https://doi.org/10.1038/nmeth.1423
Publication status	Published - Mar 2010

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abstract = "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 EF1α 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.",

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AB - 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 EF1α 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.

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MAZe: a new tool for mosaic analysis of gene function in zebrafish

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