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Printable Handouts
Navigable Slide Index
- Introduction
- Cut-and-paste DNA transposition
- The life-cycle of Tc1/mariner transposons
- Sites of molecular archeology
- Reconstruction of the Sleeping Beauty gene
- The Sleeping Beauty gene transfer system
- Delivery of Sleeping Beauty (SB)
- Hyperactive transposase mutants
- The value of intra-family sequence variation
- Hyperactive transposases transposition efficiency
- Effect of cargo size on transposition
- Mobilization of long transgenes by SB100X
- Broad applicability of SB in vertebrates
- Transposon-based technology in rodents
- Mendelian inheritance by eye
- Transposon-based technology in rats
- Transposon-based technology in rabbits
- Transposon-based technology in pigs
- Cloning of transgenic pigs by nuclear transfer
- Cloned transgenic pigs by nuclear transfer
- Transposition in human CD34+ HSCs
- GFP expression in human CD34+ HSC lineages
- A paradigm of ex vivo HSC gene therapy with SB
- Transposon-based targeted gene therapy for AMD
- Gene therapy strategy for AMD
- Stable hFIX expression in the mouse liver
- Transposons are natural mutagens
- Sequencing of transposon integration sites
- Genomic insertion patterns of integrating vectors
- Potential genotoxicity by random integration
- Minimal transcriptional activities and SB vectors
- Transcriptional shielding by HS4 insulators
- A molecular strategy for transposon targeting
- Artificial zinc finger DNA-binding domains
- Enriched SB insertions near ZF4 binding sites
- Gene transfer of iPS reprogramming factors
- iPS reprogramming of MEFs with SB
- Mouse iPS cells generated with Sleeping Beauty
- Removal of reprogramming genes
- In vitro differentiation of RMCE-d iPSCs
- Human iPS cells (hiPSCs) generated with SB
- iPSCs generated with SB express pluripotency
- Stable karyotype of hiPSCs generated with SB
- In vitro differentiation of hiPSCs
- Summary
- Acknowledgement
Topics Covered
- Transposons can be viewed as natural DNA transfer vehicles
- Transposons are capable of efficient genomic insertion, similar to integrating viruses
- Transposition can be controlled by conditionally providing the transposase enzyme
- DNA of interest cloned into a transposon-based vector can be utilized for stable genomic insertion in a regulated and highly efficient manner
- Transposition opens up many avenues for genome manipulations in vertebrates
- Generation of transgenic cells and germline-transgenic animals using transposons
- The Sleeping Beauty synthetic transposon
- Clinical applications of the Sleeping Beauty system
Links
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Talk Citation
Ivics, Z. (2014, August 5). Turning genomic junk into treasure: genetic engineering with the Sleeping Beauty transposon system [Video file]. In The Biomedical & Life Sciences Collection, Henry Stewart Talks. Retrieved October 13, 2024, from https://doi.org/10.69645/RIIV2568.Export Citation (RIS)
Publication History
Financial Disclosures
- Dr. Zoltan Ivics has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.
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
Transcript
Please wait while the transcript is being prepared...
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.
0:29
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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