Journal Article

Establishment of a gene-trap sequence tag library to generate mutant mice from embryonic stem cells

Ruiz, Patricia, Arnold, Hans-Henning, von Melchner, Harald, Otte, Jürgen, Wiles, Michael V., Lehrach, Hans, Metz, Thomas, Wurst, Wolfgang, Vauti, Franz, Füchtbauer, Annette, Füchtbauer, Ernst-Martin

2000

Overview

Nucleotide sequences of most vertebrate genes will be available in the next few years as ESTs and genomic sequences. This information will allow the deduction of encoded proteins, possibly providing clues to their functions in vivo. To determine unequivocally the physiological roles of all genes and in particular those of medical importance, mutational analyses must be performed. Large-scale random mutagenesis screens, which allow the systematic mutation of all genes, are a valuable approach to this task. To complement the sequencing efforts, we established a consortium generating a reference library of gene-trap sequence tags (GTST) which eventually will represent insertional mutations for most mammalian genes expressed in mouse embryonic stem (ES) cells. Here we report a large-scale gene-trap screen in ES cells using plasmid and retroviral gene-trap (GT) vectors that permit rapid generation and identification of mutant genes in ES cells.

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: Agriculture / Animal Genetics and Genomics / Animals / Ataxia / Biomedical and Life Sciences / Biomedicine / Breast cancer / Cancer Research / Cloning / correspondence / Embryo, Mammalian - cytology / Embryonic stem cells / Enzymes / Ewings sarcoma / Gene Function / Gene mutations / gene-trap sequence tags / Genes / Genetic aspects / Genetic Techniques / Genetic Vectors / Human Genetics / Humans / Identification and classification / Kinases / Leukemia / Mice / Mice, Mutant Strains / Nucleotide sequence / Physiological aspects / Polymerase Chain Reaction - methods / Sequence Tagged Sites / Stem cells / Stem Cells - metabolism