Description
Tol1-based transgenesis vector
The Tol1 based transgenesis vector system is a powerful DNA transposition tool for germline transgenesis of vertebrates. Tol1 is a DNA-based transposable element identified in medaka fish and demonstrated to function as an efficient gene transfer vector in a wide range of species, including mammalian cells. The system consists of a donor plasmid, containing a multicloning site flanked by terminal regions of the Tol1 element, and a helper plasmid that expresses the tol1 transposase. The transposase catalyzes the transfer of the DNA element into the host genome. The multicloning site in the donor plasmid allows ease of insertion of the gene of interest prior to delivery of both vectors into suitable germ cell line. Tol1 is capable of transferring DNA up to 22 kb, the longest cargo capacity reported for current DNA based elements. In addition, a high germline transmission frequency of 46% has been reported with tol1 vector system. Controls included in the system are helper vector expressing defective tol1 transposase and a donor plasmid containing the GFP gene cloned into the multicloning site. The Tol1 based vector system is designed to allow quick, easy transgenesis of a variety of species.