of the Max - MDC

Structure of the Group
Group Leader
Dr. Zoltán Ivics
Scientists
Dr. Oliver Walisko
Dr. Csaba Miskey
Dr. Ludivine Sinzelle
Graduate Students
Andrea Schorn
Tobias Jursch
Katrin Voigt
Ismahen Ammar
Ivana Grabundzija
Technical Assistants
Andrea Katzer
Christin Graubmann
Secretariat
Kornelia Dokup
a.
b.
c.
Figure 2. Experimental strategies for targeting
Sleeping Beauty transposition.
The common components of the targeting
systems include a transposable element
that contains the IRs (arrowheads) and a
gene of interest equipped with a suitable
promoter. The transposase (purple circle)
binds to the IRs and catalyzes transposition.
A DNA-binding protein domain (red
oval) recognizes a specific sequence
(turquoise box) in the target DNA (parallel
lines). (a) Targeting with transposase
fusion proteins. Targeting is achieved by
fusing a specific DNA-binding protein
domain to the transposase. (b) Targeting
with fusion proteins that bind the transposon
DNA. Targeting is achieved by fusing
a specific DNA-binding protein domain to
another protein (white oval) that binds to a
specific DNA sequence within the transposable
element (yellow box). In this strategy,
the transposase is not modified. (c)
Targeting with fusion proteins that
interact with the transposase. Targeting
is achieved by fusing a specific DNA-binding
protein domain to another protein (light
green oval) that interacts with the transposase.
In this strategy, neither the transposase
nor the transposon is modified.
stone in applying transposition-mediated gene delivery in
vertebrate species, including humans. We coordinate a
research project within the framework of EU FP6 with the
goal of developing novel, non-viral gene delivery technologies
for ex vivo gene-based therapies.
SB transposition occurs into chromosomes in a random
manner, which is clearly undesired for human applications
due to potential genotoxic effects associated with transposon
integration. We succeeded in targeting SB transposition
into predetermined chromosomal loci. We employed modular
targeting fusion proteins (Figure 2), in which the module
responsible for target binding can be a natural DNA-binding
protein or domain, or an artificial protein such as a designer
zinc finger. Targeted transposition could be a powerful
method for safe transgene integration in human applications.
Selected Publications
Kaufman, CD, Izsvák, Z, Katzer, A, Ivics, Z. (2005). Frog Prince
transposon-based RNAi vectors mediate efficient gene knockdown
in human cells. Journal of RNAi and Gene Silencing 1,
97-104.
Walisko, O, Izsvák, Z, Szabó, K, Kaufman, CD, Herold, S, Ivics, Z.
(2006). Sleeping Beauty transposase modulates cell-cycle progression
through interaction with Miz-1. Proc. Natl. Acad. Sci.
USA 103, 4062-4067.
Ivics, Z, Izsvák, Z. (2006). Transposons for gene therapy! Curr.
Gene Ther. 6, 593-607.
Ivics, Z, Katzer, A, Stüwe, EE, Fiedler, D, Knespel, S, Izsvák, Z.
(2007). Targeted Sleeping Beauty transposition in human cells.
Mol. Ther. 15, 1137-1144.
Miskey, C, Papp, B, Mátés, L, Sinzelle, L, Keller, H, Izsvák, Z,
Ivics, Z. (2007). The ancient mariner sails again: Transposition
of the human Hsmar1 element by a reconstructed transposase
and activities of the SETMAR protein on transposon ends. Mol.
Cell. Biol. 27, 4589-600.
108 Cancer Research