CATALYSTS
‣ Today , modern research in molecular biology uses different tools for genome editing . What they all have in common is that they work on the basis of nuclease enzymes that specifically recognise a gene locus and do so by interacting DNA-binding domains of the nuclease ( s ) with a sequence of bases in a predetermined genome section that is as unique as possible . This class of nucleases , which usually induce cleavage of bound DNA , includes zinc-finger nucleases ( ZFNs ), transcription activator-like effector nucleases ( TALENs ) or meganucleases ( Figure 1 ). In the early 20th century , clustered , regularly interspaced , short palindromic repeat ( CRISPR ) -Cas systems were discovered and with them systems that function differently . In CRISPR systems , a specific RNA mediates the recognition and targeting of DNA . With the discovery of * CRISPR- Cas systems ( Cas9 , Cas12 or others ) and their ability to modify DNA precisely , a tool had been discovered that , by following nature , revolutionised the biotechnologicallyinduced evolutionary process of living organisms . With the emerging technology , which was first applied in 2012 , the selection process could not only be carried out in a targeted and precise manner , it could also be accelerated enormously . Biotechnology can therefore now selectively insert , remove or modify individual DNA segments in living organisms in a relatively simple process . Some people are already talking about so-called garage technology for programming microorganisms that can be used industrially . An ingenious technology – were it not for the dispute over patents , which makes the current legal situation for commercial use complex and uncertain and also demands high licensing fees from companies for using the technology . The uncertain legal situation and the licensing agreements for the commercial use of CRISPR technology , which are financially unsustainable for many companies , have motivated us to develop our own variant of the CRISPR-Cas scissors with the BRAIN- Engineered-Cas ( BEC ) nuclease . This enables us , for example , to optimise the metabolic performance of microbial production strains within a short time . In view of this licensing situation , we turned to our technological expertise and set ourselves the goal of developing our own nuclease . We used metagenome sequencing and protein engineering techniques to identify a Class 2 CRISPR-associated nuclease that has low sequence homology to other CRISPR nucleases and targets DNA with a unique molecular mechanism . We have filed a patent to protect the nuclease DNA sequence and expect to be able to use this system freely in the future .
‘ We CRISPR for you ’
We not only use our validated CRISPRassociated BEC nuclease for targeted gene editing in our own projects , but also make it available for customer projects . Organisms in which BEC targets DNA include bacteria , fungi and yeasts . Accordingly , the technology is being used in these organisms for collaborative projects and is also being progressively validated for industrial use . Activity in plants has also been demonstrated , but validation is pending . Genome editing tests for other applications , such as mammalian cell lines , are currently underway . But what exactly does the use of this nuclease mean for users ? The technology is an enabling technology and allows the user to precisely optimise microorganisms according to his own specifications . Once established in the target organism , genome editing is not only precise , but also efficient and fast . It is often superior to conventional technologies in terms of time to market : the development of , for example , highly efficient producer strains can be significantly accelerated . Further potential applications of BEC technology lie in the food
© BRAIN Biotech up the side
industry . In some countries , genome editing is already applied in food manufacturing processes , including precision fermentations to obtain alternative proteins from microbial or plant sources or optimisation of microbial strains to produce fermented beverages . In order to drive sustainability in companies , the chemical industry can switch part of the chemical processes to biological processes . Here , for example , BEC nuclease can modify microbial strains to use a specific organic substrate as a carbon source and to engage in synthesis pathways in the course of which the microorganisms produce bio-based building blocks and platform chemicals . Converting
34 SPECIALITY CHEMICALS MAGAZINE ESTABLISHED 1981