Transgenerational analysis of transcriptional silencing in zebrafish.

摘要: The yeast Gal4/UAS transcriptional activation system is a powerful tool for regulating gene expression in Drosophila and has been increasing popularity developmental studies zebrafish. It also useful studying the basis of de novo silencing. Fluorescent reporter genes under control multiple tandem copies upstream activator sequence (UAS) often show evidence variegated DNA methylation transgenic zebrafish embryos. To characterize this systematically, we monitored progression silencing UAS-regulated transgenes that differ their integration sites repetitive nature UAS. Transgenic larvae were examined three generations tissue-specific green fluorescent protein (GFP) at Single insertions containing four distinct sequences far less susceptible to than fourteen same In addition, integrated or adjacent transposon exhibited regardless number UAS included transgene. Placement promoter-driven Gal4 responder single bicistronic construct appeared accelerate methylation. results demonstrate utility efficient tracking mechanisms across several generations, as well provide guidelines optimal Gal4-regulated organisms subject