Turning genomic junk into treasure: genetic engineering with the Sleeping Beauty transposon system

Published on August 5, 2014   59 min

A selection of talks on Infectious Diseases

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0:00
My name is Zoltan Ivics. I work at the Division of Medical Biotechnology at the Paul Ehrlich Institute in Langen, Germany. And the subject of this presentation will be how we turn genomic junk into treasure, an introduction to transposable elements, and how we use these transposons, namely the Sleeping Beauty system, for genetic engineering in animals, and for molecular therapy.
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Just to introduce to you how transposons work I'd like to outline the so-called Cut-and-paste DNA transposition mechanism. The transposable element, or transposon in short, is depicted here on the top of this slide. The very ends of the transposon are indicated as black arrows. These are the so-called terminal inverted repeats of the transposon. These sequence are important for the transpositional reaction. And these repeats flank in the natural context a gene that encodes the transposase protein, which is the enzymatic factor of the transpositional process. So for transposition to occur the transposase needs to be expressed, followed by sequence specific binding of the transposase molecules to the very ends of the transposon. And then this is followed by a so-called synaptic complex formation, in which the two ends of the transposable elements are grouped together by transposase interactions. And then this step is followed by physically removal of the transposon out of its original DNA context. This is called transposon excision. And then this excised transposon will interact with a new piece of DNA that is highlighted here by the green color. And the transposon will integrate into this new DNA, giving rise to a relocated or transposed piece of DNA. So transposition now occurs from the yellow DNA into the green DNA, and then the transpositional reaction, the excisions that generates damage, double stranded DNA breaks at the ends of the transposon, which will be repaired by host DNA repair mechanisms.

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Turning genomic junk into treasure: genetic engineering with the Sleeping Beauty transposon system

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