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With Molecular "Green Thumb"

Live in the Lab: Genome Editing as a New Tool for Plant Breeding

| Author: Dr. Ilka Ottleben

It doesn't work without a team

"With us, not everyone works rigidly on their project, but everyone helps each other," emphasizes Sprink. The two TAs Annett Beyer and Jana Goldschmidt are responsible for all maintenance work in the laboratory. For example, they produce plant media and maintain the tissue cultures. If time is short with other employees, Annett Beyer also takes care of plant transformations. She is also Sprink's number one contact if there are short-term or new issues to be addressed, e.g. leading to new projects. "Annett's the one who's gonna push it and do the work first." This requires knowledge of all cultures and techniques. "The varied tasks with constantly new methods and the work in our young, dynamic team are fun and motivating for me," says Beyer. Indeed, there are many methods that need to be mastered.

"We are a typically molecular biological laboratory, from DNA and RNA isolation to PCR, qPCR and all common cloning techniques to protein purification, we cover all common techniques," explains Sprink. "We try to use as few toxic chemicals as possible, such as ethidium bromide or chloroform. Wherever possible, we work with alternative substances or switch to kits that contain less toxic substances." In addition, there are all common plant transformation techniques.

"If you're working on off-targets, you also need to know the genome very well. We therefore do a lot of sequencing of often very long sequences. On average we get 20 to 30 sequencings per week. For this, we use not only Sanger but also SMRT sequencing. The sequencing itself has been outsourced, and we have excellent support from bioinformatics in the evaluation of the sequences."

On the equipment side, the employees currently feel well equipped, even though "air is always up" in principle. The purchase of large equipment worth 5000 euros must be applied for at the JKI two years in advance or is financed by the institutes via overhead funds. For example, a new fluorescence microscope has recently been purchased to help analyze DNA-free transformed plant cells, and this year they are investing in new culture cabinets to increase capacity and improve culture management. Moreover, the number of incubation shakers for agrobacteria and E.coli cultures is sometimes a bottleneck in the AG, whereby a new device was added last year. "Space is always an issue," says Janina Metje-Sprink. Time is also a factor: the purchase of a digital PCR device is planned for this year. For special applications such as FACS (Fluorescence Activated Cell Sorting) or micromanipulation, the AG cooperates with other institutes or tries to initiate such cooperations if necessary.

Genome editing: More opportunities or risks?

The Sprink group has been working together for a good 18 months to investigate genome editing, its applications and possible risks. Against the background of the effort involved, this time is not so long, but is it sufficient to make initial statements? "So far, we have not identified any risks, but we are continuing our research," says Sprink. "If off-targets occur, this usually seems to be not due to the technology, but rather the user. In addition, we speak of few off-targets, compared to mutagenesis methods, such as irridation, which are common today. Those methods produce several thousand off-targets, which also occur completely undirected. So you always have to compare them," he continues.

"Of course, applications are also possible with the GE that clearly lead to the genetically-modified organism and thus should and will be regulated as such. On the other hand, other applications simply add very specific mutations that can also occur naturally. I am very strongly in favor of looking at this in a differentiated way," sums up Sprink. Against this background, the researcher is disappointed by the current ruling of the European Court of Justice, which confirmed in summer 2018 that all organisms obtained by mutagenesis must be regulated explicitly according to the current GMO Directive.

"From our ElsaGEA project, which aims to compile ethical, legal and socio-economic aspects of genome editing in agriculture as comprehensively and systematically as possible, we also know that, contrary to some speculation, only about 14% of GE applications have herbicide tolerance as their sole breeding objective. On the other hand, the majority of applications target biotic and abiotic characteristics, such as increased yields, improved food quality or higher tolerance to diseases, pests and abiotic stress such as those caused by climate change. Breeding already has an enormous range of tools at its disposal. Genome editing is another tool that has many advantages, but certainly not everything is possible with it. But we should seize the opportunity to democratize technology so that many breeders - even small ones - can benefit from it."

* Dr. I. Ottleben Editor LAB Worldwide/LABORPRAXIS, E-Mail ilka.ottleben@vogel.de

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