There are currently no consolidated systems that can both upregulate and downregulate the translation of specific messenger RNA (mRNA) targets. Known methods to achieve targeted downregulation include anti-sense oligonucleotides (ASO) and short interfering RNAs (siRNA). However, both of these technologies function to destabilize a messenger RNA target and downregulate translation, rather than upregulate translation. There are few known methods to increase mRNA translation and these methods are not well characterized.
As such, there is a need to provide compositions and methods for recruiting translational pre-initation complexes in trans and thereby control translation in cells and in gene therapy techniques. Currently, there are no consolidated systems that can both upregulate and downregulate the translation of specific messenger RNA targets.
Researchers from UC San Diego have developed a technology to control translation in cells using CRISPR-Cas protein fusions. The inventors refer to this technology as Controlled Translation via Protein effectors (ConTra). This technology is based on the 5’Cap binding biology of EIF4E protein and its regulator EIF4E-BP1, which enhance and repress translation, respectively.
This patent-pending technology relates to compositions, systems, methods, and kits to control mRNA translation in cells using CRISPR-Cas protein fusions. These compositions, methods, systems, and kits utilize the RNA targeting abilities of CRISPR-Cas systems, which use a guide RNA to provide a simple and rapidly programmable system for recognizing RNA molecules in cells. These compositions, methods, systems, and kits further utilize the ability of CRISPR-Cas systems to bind target messenger RNA to initiate translation in trans by fusing a ribonucleic acid sequence, that recruits translational pre-initiation complexes, to the single stranded guide RNA and thereby to the bound messenger RNA. CRISPR-Cas systems also have neutral effects on messenger RNA stability, which makes any measured change to protein expression a function of the fused protein effector. The compositions, systems, methods, and kits described herein provide high utility and versatility when compared to other compositions, methods, systems, and kits for controlling mRNA expression.
Applications for these methods include the use as a research tool for controlled translation of a messenger RNA. Additionally, it can be used in therapeutics whereby Viral (AAV) or other delivery approaches of ConTra to treat diseases rooted in haploinsufficiency.
This application can be extended to other CRISPR-Cas systems such as other Cas9 orthologs, Cas13a/b orthologs, CasRX/Cas13d orthologs that target RNA. Any messenger RNA of interest can be targeted, given the selection of an appropriate spacer sequence, which is specific to each CRISPR-Cas system.
UC San Diego is seeking partners to commercialize this patent-pending technology.
|Patent Cooperation Treaty||Published Application||WO2019204828||10/24/2019||2018-296|
CRISPR-Cas, cellular RNA, mRNA translation, post-transregulation of gene expression