In order to produce designer organisms for a variety of commercial, diagnostic, and therapeutic uses, synthetic biology aims to achieve robust control of biological processes. Researchers at the iNANO center of Aarhus University have made RNA origami sponges and CRISPR-based regulators for advanced genetic control of enzymatic pathways in microorganisms. They did this to make it easier to make valuable biochemicals.
One of the main tenets of the now-mature field of synthetic biology has been the development of tools for precise control of biological processes. These scientific tools incorporate ideas from numerous research areas, resulting in novel applications that may have a profound impact on contemporary society. Due to compatibility with folding and expression in cells, translating modern RNA nanotechnology innovations into the biological context has enormous potential. However, it also presents special challenges, such as stringent performance requirements and the inherent instability of RNA molecules.
The Andersen lab recently developed a structural RNA design method called “RNA origami” that aims to address this. The goal of this strategy is to produce sophisticated RNA-based devices made by humans that are stable in cells, interact with other biomolecules like proteins and other RNA, and support novel uses, particularly in the context of gene regulation. RNA origami is presented as a sophisticated RNA design platform that, when used in the context of a cell, creates unique molecules for synthetic biology-based regulation. This is shown by two different approaches that were just published.
RNA sponges regulate enzyme production in bacteria
The first method, when expressed in bacteria, the RNA origami, was used to achieve precise control of protein production levels. By adding a potent protein binding site to the expressed protein’s own gene, self-inhibiting protein expression cassettes were created. The same protein-binding sites were then expressed in large quantities in RNA origami. As a result, the self-inhibited protein can be expressed because the RNA origami acts as a protein sponge to bind proteins inside the cell. This general idea has been shown to make it possible to control a lot of proteins at once and turn on enzymatic pathways to make more products.
CRISPR-based regulators for yeast chemical factories
The second method controls gene expression in yeast by combining RNA origami with CRISPR, one of the most well-liked contemporary molecular biology techniques. The small RNAs that direct CRISPR-Cas9 to target particular sequences in the DNA genome have the RNA origami integrated into them. In order to attract transcription factors, protein-binding sites were added to the RNA origami scaffolds. The transcription factors induced gene expression by concentrating on promoter regions and binding the RNA scaffolds there. It has been demonstrated that the scaffold’s orientation and the number of transcription factors it recruits can be used to adjust the expression strength. Finally, it was shown that it was possible to control multiple enzyme pathways for high-yield violacein production.
