Ntransposon-mediated bac transgenesis in zebra fish books

It avoids potential multicopy concatameric integrations, allowing confirmation of the completeness of the bac from end to end, as well as mapping of the genomic integration site. Pdf transposonmediated bac transgenesis in zebrafish. The zebrafish information network zfin is the database of genetic and genomic data for the zebrafish danio rerio as a model organism. Transposonmediated bac transgenesis is a new tool for precise delivery of bacs in both zebrafish and mice. Identification and characterization of alternative promoters of zebrafish rtn4nogo genes in cultured cells and zebrafish embryos. We took advantage of the precise cutandpaste behavior of a natural transposon, tol2, to develop a new method for bac transgenesis. Overexpression of bim and bid mediates apoptosis in asxl1. Bacterial artificial chromosomes bacs are among the most widely used tools for studies of gene regulation and function in model vertebrates.

Here we report a stepbystep protocol for efficient bac transgenesis in zebrafish using the medaka tol2 transposon. A plasmid set for efficient bacterial artificial chromosome bac. Nitroreductasemediated cell ablation in transgenic zebrafish. In contrast, crisprcas9mediated genome editing technology now provides a great opportunity. The most popular system for generating transgenic animals in zebrafish is the tol2. Cremediated recombination allows the induction of protein expression in a. Transposonmediated bac transgenesis in zebrafish nature. Nitroreductasemediated cell ablation in transgenic zebrafish embryos. Identification and characterization of alternative promoters of zebrafish rtn. Transposonmediated bac transgenesis in zebrafish and mice. Background bacterial artificial chromosomes bacs are among the most widely used tools for studies of gene regulation and function in model vertebrates, yet methods for predictable delivery of bac. Ijms free fulltext zebrafish models of neurodevelopmental. Rapid bac selection for tol2mediated transgenesis in zebrafish. Recently, tol2 transposasemediated transgenesis has emerged as a.

Despite being a nonmammalian animal, the zebrafish model has proven its potential in. Transgenesis in zebrafish with the tol2 transposon system. Zebrafish danio rerio is a fresh water fish that inhabits rivers in india. The tol2bac was coinjected with the transposase mrna into zebrafish or mouse eggs. Cold tolerance ability in the transgenic zebrafish tg smyd1. Analyzing cold tolerance mechanism in transgenic zebrafish. Transposonmediated bac transgenesis in zebrafish and mice article pdf available in bmc genomics 101. Search for mutants, knockdowns, transgenics, and affected phenotypes. Bac transgenesis has been used extensively in a broad range of applications in mice, from studies of gene regulation to creating animal models of human disease 1,2,3. A guide for the laboratory use of zebrafish danio brachydanio rerio.

Transposonmediated bac transgenesis in human es cells transposonmediated bac transgenesis in human es cells. The tol2bac portion was excised in vivo and integrates as a single. Using recombineering in escherichia coli, we introduce the itol2 cassette in. In this new method, the minimal sequences of the tol2 transposon were used to deliver precisely single copies of a 70 kb bac transgene to the zebrafish and mouse genomes.