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  • A Multifunctional Mutagenesis System for Analysis of Gene Function in Zebrafish.

A Multifunctional Mutagenesis System for Analysis of Gene Function in Zebrafish.

G3 (Bethesda, Md.) (2015-04-04)
Helen Ngoc Bao Quach, Shijie Tao, Pavle Vrljicak, Adita Joshi, Hua Ruan, Rashmi Sukumaran, Gaurav K Varshney, Matthew C LaFave, Shawn M Burgess, Christoph Winkler, Alexander Emelyanov, Sergey Parinov, Karuna Sampath
ABSTRACT

Since the sequencing of the human reference genome, many human disease-related genes have been discovered. However, understanding the functions of all the genes in the genome remains a challenge. The biological activities of these genes are usually investigated in model organisms such as mice and zebrafish. Large-scale mutagenesis screens to generate disruptive mutations are useful for identifying and understanding the activities of genes. Here, we report a multifunctional mutagenesis system in zebrafish using the maize Ds transposon. Integration of the Ds transposable element containing an mCherry reporter for protein trap events and an EGFP reporter for enhancer trap events produced a collection of transgenic lines marking distinct cell and tissue types, and mutagenized genes in the zebrafish genome by trapping and prematurely terminating endogenous protein coding sequences. We obtained 642 zebrafish lines with dynamic reporter gene expression. The characterized fish lines with specific expression patterns will be made available through the European Zebrafish Resource Center (EZRC), and a database of reporter expression is available online (http://fishtrap.warwick.ac.uk/). Our approach complements other efforts using zebrafish to facilitate functional genomic studies in this model of human development and disease.

MATERIALS
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