| A simple and highly efficient transgenesis method and a new method for BAC transgenesis in mice with the Tol2 transposon system | ||
| Genomics Feb/26/2010（Epub）/ BMC Genomics Oct/16/2009（Epub） | ||
| Saitou Laboratory, Division of Population Genetics Kawakami Laboratory, Division of Molecular and Developmental Biology |
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| 1) A simple and highly efficient transgenesis method in mice with the Tol2 transposon system and cytoplasmic microinjection. Sumiyama, K., Kawakami, K., and Yagita, K. Genomics 95(5) 2010. doi:10.1016/j.ygeno.2010.02.006 2) Transposon-mediated BAC transgenesis in zebrafish and mice. Suster, M.L., Sumiyama, K., and Kawakami, K. BMC Genomics 10, 477, 2009. doi:10.1186/1471-2164-10-477 Transgenesis is an essential technology for genetic studies. Although transgenic mice are currently created by pronuclear injection of plasmid DNA, the efficiency is low, i.e., only ~3% of the injected eggs give rise to germline transmitting founders. In collaboration of two labs in NIG, a novel transgenesis method has been developed, which increased the transgenic frequency drastically. In this method, a foreign DNA cloned in a Tol2 -transposon vector is injected with the transposase mRNA into the cytoplasm of fertilized eggs. Both survival of the injected embryos and integration of the foreign DNA into the genome are enhanced, and, consequently, the overall transgenic efficiency becomes constantly more than 20%. BAC transgenesis is an important technique to study the function of an entire gene or cis-regulatory elements. However this technique is hindered by low transgenic frequencies, concatemeric integrations and unwanted rearrangements of the target sites. To circumvent these problems, we have shown that a Tol2 vector can carry a DNA insert of a BAC size and developed a new method for BAC transgenesis by using the Tol2 transposon system. We expect these methods will revolutionize mouse transgenesis and should facilitate high-throughput functional genomics studies.
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