2.<I>Sleeping Beauty (SB) </I>トランスポゾンシステムによるマウス遺伝子の大規模スクリーニング戦略
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概要
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Generation of mutant mice with the conventional gene-targeting method is useful approach in reverse genetics. However, the desired phenotype may not be observed easily because this approach is labor-intensive and the only one gene can be mutated at a time. In contrast, phenotype-driven forward genetics provide great advantages to analyze the mutated functional genes. Although, large-scale production of mutant mice by chemical mutagen has been reported, detection of induced point mutation responsible for phenotype is difficult and time-consuming. The <I>Sleeping Beauty (SB) </I> transposon is a mobile DNA element jumping from one location to another, and has been developed as a tool for insertional mutagenesis. Determination of the insertion site is facilitated by using the transposon sequence as a molecular tag. The efficient activity of the <I>SB</I> transposase in the mouse germline provides us with a useful system for random germline insertional mutagenesis. We have tested <I>SB</I> transposon vector for use in mouse germline insertional mutagenesis. The offsprings generated from mating transgenic males harboring transposon and transposase with wild-type females demonstrated efficient transposition in mouse germline. The newly developed transposon vector combined with gene trap method and GFP reporter enabled us to select for mutant mice rapidly and non-invasively. The homozygous mutant mice generated with this vector showed apparent phenotype, indicating that our transposon vector was highly mutagenic. Taken together, these results indicate that the SB transposon system has promise as a new powerful tools for large-scale genetic screening in mice.
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