Click-SELEX - Novel method for generation of selection from a vast spectrum of functional oligonucleotides
Keywords: Click-chemistriy, orthogonal chemistry, SELEX, aptamer, functional oligonucleotides, libraries, Screen, pharma, red biotech, White biotech, molecular biology, chemical, therapeutic, modular, DNA, Uni Bonn
Here, in an iterative process, a pool of single stranded oligonucleotides is contacted with a target structure (e.g. a protein), high-affinity oligonucleotides are then isolated, amplified and again put into contact with the target until, eventually, an ideal oligonucleotide has been obtained. SELEX has opened up new avenues e.g. for generating aptamers as therapeutic compounds - and thereby enlarging the space for novel lead structures. However, the chemical space and thus the number of targetable proteins is still restricted by the limited diversity of nucleic acids. Use of chemically modified nucleic acids can solve this issue, but such nucleotides often lack compatibility with enzymatic steps (e.g. PCR) of the in vitro selection process. Here, a versatile method (Click-SELEX) has been developed for the modular expansion of the chemical space of nucleic acid libraries, thus enabling the generation of nucleobase-modified aptamers with unprecedented recognition properties. The introduction of additional chemical entities into the DNA is achieved by using a 5’ modified, commercially available nucleoside-triphosphate whereas the modification can be further derivatized with so-called bio-orthogonal chemistry, e.g. click-chemistry. Advantageously, the modification is only transiently present during the selection cycle. This enable a high degree of compatibility with the enzymatic step of the selection process. This approach further allows introducing larger chemical entities into DNA libraries that cannot be used using current methods. The click chemistry has a broad acceptance of diverse substrates, which are either commercially available or accessible by chemical synthesis.
- Access to a vast spectrum of functional oligonucleotides with unprecedented properties
- Simple, robust, efficient protocol
On behalf of University of Bonn, PROvendis offers licenses for commercial use as well a research collaboration with licensing option.
An International and a European patent application are pending:
International Patent Application WO 2015/049359
International Patent Application WO 2016/050850
Mayer, G., et al. (2016) Click reaction on solid phase enables high fidelity synthesis of nucleobase-modified DNA. Bioconjug. Chem. Feb 5., epub ahead of print.
Dellafiore, M.A., et al. (2016) Modified nucleoside triphosphates for in-vitro selection techniques. Front. Chem. 4: 19, epub.
Tolle, F., et al. (2015) A versatile approach towards nucleobase-modified aptamers. Angew. Chem. Int. Ed. Engl. 54(37): 10971-4.
Eine Erfindung der Uni Bonn.