of the Max - MDC

a.
b.
Figure 1. The Sleeping Beauty transposon system. (a) Components and structure of a two-component gene transfer system based on
Sleeping Beauty. A gene of interest (orange box) to be mobilized is cloned between the terminal inverted repeats (IR/DR, black arrows)
that contain binding sites for the transposase (white arrows). The transposase gene (purple box) is physically separated from the IR/DRs,
and is expressed in cells from a suitable promoter (black arrow). The transposase consists of an N-terminal DNA-binding domain, a nuclear
localization signal (NLS) and a catalytic domain characterized by the DDE signature. (b) Mechanism of Sleeping Beauty transposition.
The transposable element carrying a gene of interest (GOI, orange box) is maintained and delivered as part of a DNA vector (blue DNA). The
transposase (purple circle) binds to its sites within the transposon inverted repeats (black arrows). Excision takes place in a synaptic complex.
Excision separates the transposon from the donor DNA, and the double-strand DNA breaks that are generated during this process are
repaired by host factors. The excised element integrates into a TA site in the target DNA (green DNA) that will be duplicated and will be flanking
the newly integrated transposon.
zebrafish. Transposition of SB is enhanced by CpG methylation
of transposon donor DNA, and we are currently testing
models for the enhancing effect of DNA methylation.
Loss-of-function insertional mutagenesis
Ivana Grabundzija
Transposons can be applied as useful research tools for gene
discovery, thereby contributing to our understanding of
gene function in vertebrates. We are taking advantage of
local hopping for regional saturation mutagenesis in mice,
where the primary transposon donor locus can be determined
by targeting the transposon to a chromosomal region
of interest. We began to work on regional transposon mutagenesis
of the Williams-Beuren syndrome locus (in collaboration
with Thomas Floss, GSF), with the goal to uncover the
genetic basis of this disease.
Transposons as non-viral vectors for gene
therapeutic approaches
Ismahen Ammar, Csaba Miskey, Katrin Voigt
DNA-based transposons are natural gene delivery vehicles,
and molecular reconstruction of SB represents a corner-
Cancer Research 107