AgroJector is a plant biotechnology company striving to inject enabling, precise, biosafe, affordable and environment friendly
technologies and research supplies for plant research community.
AgroJector shall significantly contribute to the bio-economy, help tackling the challenges arising from human population growth and climate change and foster technologies increasing productivity and sustainability but reducing environmental pollution.
Plant scientists and agriculture biotechnology companies face
dificult tasks to address the needs of human population growth,
adaptation to climate change and environment pollutions. They are
expected to generate higher yielding crops, increase plant
tolerance to cold, drought, pathogens and pests, as well as find
ways of reducing the use of environment polluting chemicals
in agriculture. Similarly they should support booming alternative
industries that make use of plants and plant products in
biofuel, bioplastic, building, clothing and medical industries.
Finally, the solutions developed should be safe and environment
AgroJector will develop enabling tools and technologies with an unmatched specificity and biosafety for the introduction of lost plant traits into modern crops as well as non-transgenic means of molecular breeding of crops. The indispensable core products for these technologies are patent-pending biosafe Agrobacterium strains and associated cloning and expressing vectors currently being developed by the company.
Agrobacterium tumefaciens naturally delivers proteins and DNA to plants. The simple and highly efficient trans-formation procedure working on a broad range of plant species made it the most commonly used tool for plant transformation in research and industry for the last 20 years.
All currently used strains of A. tumefaciens have an important biosafety problem as they uncontrollably transfer large pieces of genomic DNA to plants (Ulker et al. 2008). This makes it nearly impossible to guarantee the exclusive transfer of proteins and thereby the highly specific changes of plant genomes.
The use of the same modern gene targeting proteins that are under investigation for human gene therapy makes highly specific, minimal changes in plant genomes possible (Fichtner et al., 2014). The resulting difference in sequence is less than between any two individual plants.
After five years of research, the mechanism for the transfer of genomic DNA could be unraveled and provided the basis for the first biosafe Agrobacterium strains, AgroSafe. Four distinct genomic regions that were found to cause the transfer were deleted in the strains. The patent pending strains are the indispensible basis for various novel tools.
Genetic modification is the most important tool in
understanding how plants grow and function. The
method of choice for this purpose is Agrobacterium
tumefaciens. Agrobacterium is capable of transfering
any desired sequence of interest flanked by sequences
called left border (LB) and right border (RB) - altogether
called T-DNA - to plants. For this purpose, the T-DNA is
cut and funneled by VirD2 through the natural TypeIV
secretion system (Type IV SS).
Specifitiy of gene delivery is crucial for this technology. For more than 20 years, scientists thought that Agrobacterium exclusively transfers the sequence of interest to plants. However, it was shown that Agrobacterium also uncontrollably transfers large pieces of genomic DNA (Ulker et al. 2008) which significantly compromises the necessary specificity and biosafety. AgroJector develops the first biosafe Agrobacterium strains AgroSafe by deleting all elements that were found to activate the transfer of genomic DNA.
1, 2, 3-VirD2 relaxase (M, DONOR DNA binder) from Agrobacterium
tumefaciens is fused to a TAL type programmable DNA binding protein
(W, ACCEPTOR DNA binding protein) with a linker region (L) allowing
flexibility of M and -W. VirD2 relaxase can also be fused to Cas9 protein.
4-Any DNA cloned between RB-LB in T-DNA can be transferred and targeted to a specific locus in plant genome (5).
6-In order to increase the efficiency of gene targeting, a DNA cleavage approach at the target site can be undertaken by fusing DNA targeting TAL factors to molecular scissors such as FokI nuclease.
M and W can be TAL, Cas9 or Zink finger type DNA binding proteins.
Single base pair changes
Target desired genomic DNA regions in plants with unmatched specificity.
Brings your DNA fragment closer than 100°A to your target.
Any DNA cloned between RB-LB in T-DNA can be transferred.
AgroSAFE Agrobacterium strains are necessary.
Biosafe reintroduction of lost genes in natural loci (re-wilderung).
Native origin of the Ti plasmid for T-DNA plasmid leads to reduced copy number of T-DNA. Thereby reducing silencing in plants due to lower number of insertion sites
Deleted origin of transfer led to the unintended transfer of up to 50kb chromosomal DNA with a frequency of about 10%.
Deleted transposons were shown to interrupt virulent genes, activate antibiotic resistance gene and jump into the T-DNA, being transferred to plants
Integration of the virulent genes of the Ti plasmid into the genome reduces necessary antibiotics by one.
single base pair changes
SeedSELECT requires much less space than convential selection of plants. Thereby, the cheap and simple generation of genetically modified plants in a high-throughput fashion is made possible.
Due to the easy and rapid selection of genetically modified plants, transformation protocols using Agrobacterium can be unprecedentedly rapid optimised in order to increase the host range.
of plants needed
Six of the eight co-founders of AgroJector are experienced plant scientists driven by their passion for the development of precise and biosafe technologies.
Dr. Bekir Ülker (CEO), a former group leader at the University of Bonn and Dr. Franziska Turck (Scientific Advisor), a research group leader at the Max- Planck
Institute for Plant Breeding Research, both have more than 20 years experience in plant molecular biology research.
In 2008, Dr. Ülker and his collaborating researchers were the first to discover the unexpected biosafety risk in publicly available Agrobacterium strains. Five subsequent years of DFG funded pioneering research in Dr. Ülkers former group were necessary to unravel the mechanism of this undesired transfer. Now knowing the bases of the undesired transfer, prototypes obsolete of most of the undesired transfer were developed. This was the prerequisite for the patent pending, first available biosafe Agrobacterium strains AgroSafe.