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Technologies

A core technology of the MDCell is a novel Sleeping Beauty (SB) transposon system.

It is a DNA/RNA-based gene engineering tool that does not require the generation of viral vectors. Thus, it provides a great opportunity to facilitate cell engineering for research and clinical use.

The SB system consists of 2 components where a transposase enzyme medicates the integration of a transposon genetic element into a target genome. To do so, the transposase binds to inverted repeats (IR) of the transposon vector that flank the transgene cassette and paste it into the genome of a host cell. It has been found that this process is random and SB has no preference to integrate in transcriptionally active or oncogenic gene regions.

To generate a SB system that provides high transfection rates upon a single modification step while maintaining high cell viability, three technical advances have been achieved:

  1. A hyperactive transposase (SB100X) with increased transposition efficiency has been developed at the MDC in the research group of Dr. Zsuzsanna Izsvák.
  2. To complement the SB100X transposase a novel transposon vector (pT4) has been developed by molecular evolution at the MDC in the research group of Dr. Zsuzsanna Izsvák.
  3. For a long time, broad application of DNA transfection has been hindered by the toxic effect that DNA transfection has on cells. Especially for primary cells, DNA transfection negatively influences cell viability. To overcome this problem the research group of Prof. Dr. Wolfgang Uckert developed a technology to apply the SB system in primary human T cells. This enables stable transfection rates of TCRs or CARs of 50% upon a single modification step while maintaining a high survival rate and full functionality of the T cells.

Gene and cell therapy products for clinical application require complex manufacturing processes. For clinical development it is necessary to already pick the best suited gene engineering system early on during discovery and pre-clinical phase, because changes in the manufacturing process during clinical development is an intricate procedure. For pre-clinical development it is also important to consider a gene engineering system where licenses for drug development are available.

Thus, pick MDCell’s novel Sleeping Beauty transposon system or MDCell’s universal retrovirus gene engineering platforms  for feasible and reliable gene insertion at high efficiency.

Advantages of MDCell’s Sleeping Beauty transposon system for clinical use in TCR / CAR T cell therapy

 

Sleeping Beauty

Viral vectors

CRISPR/Cas9

Knock-in efficiency

High

High

Low

Genome integration pattern

Safe (random)

Safe (biased)

Safe (targeted)

Time of vector production

Short

Long

Short

Costs of GMP production

$

$$$

$$

Biosafety level

S1

S2

S1

Cargo size

Large

Medium

Large

Freedom to operate (FTO)

MDCell technology

MDCell technology

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