Mammalian mutagenesis using a highly mobile somatic Sleeping Beauty transposon system

Abstract

Transposons have provided important genetic tools for functional genomic screens in lower eukaryotes but have proven less useful in higher eukaryotes because of their low transposition frequency. Here we show that Sleeping Beauty (SB), a member of the Tc1/mariner class of transposons, can be mobilized in mouse somatic cells at frequencies high enough to induce embryonic death and cancer in wild-type mice. Tumours are aggressive, with some animals developing two or even three different types of cancer within a few months of birth. The tumours result from SB insertional mutagenesis of cancer genes, thus facilitating the identification of genes and pathways that induce disease. SB transposition can easily be controlled to mutagenize any target tissue and can therefore, in principle, be used to induce many of the cancers affecting humans, including those for which little is known about the aetiology. The uses of SB are also not restricted to the mouse and could potentially be used for forward genetic screens in any higher eukaryote in which transgenesis is possible.

A flk‐1 promoter/enhancer reporter transgenic Xenopus laevis generated using the Sleeping Beauty transposon system: An in vivo model for vascular studies

“…It has also been used as a gene therapy vector in the treatment of mouse models of human genetic diseases (Hackett et al, 2005). Insertional mutagenesis screens using SB have been carried out in mice and zebrafish and have identified genes involved in cancer as well as early development (Collier et al, 2005;Dupuy et al, 2005;Sivasubbu et al, 2006).…”

Chromosomal mobilization and reintegration of Sleeping Beauty and PiggyBac transposons

“…SB has proven to be particularly effective as a somatic mutagen to identify oncogenes by insertional mutagenesis (Collier et al, 2005;Dupuy et al, 2005). SB has also been used for germ line mutagenesis but has been less effective in this context (Dupuy et al, 2001;Fischer et al, 2001;Horie et al, 2001).…”

Generation and Characterization of a Transgenic Zebrafish Expressing the Reverse Tetracycline Transactivator

“…Since then, many commercial fish species such as carp, tilapia and salmonids have been successfully used for transgenesis (Martinez et al, 1996;Cook et al, 2000;Sun et al, 2005). Foreign genes are usually introduced into a target genome by microinjection, electroporation, sperm, transposon and retrovirus-mediation (Kawakami et al, 2000;Nagano et al, 2001;Dupuy et al, 2005). In addition to commercial transgenic fish, transgenic models of zebrafish and medaka have been developed in many laboratories in order to understand the mechanisms of growth, embryonic development, disease resistance, and aspects of certain human diseases (Wittbrodt et al, 2002;Feitsma and Cuppen, 2008).…”