Background

Nonsense mutations, or premature termination codons (PTCs), make up 10-15% of all genetic diseases, affecting over 30 million people worldwide through 7,000 different diseases. Currently, there are very limited options for their therapeutic management. Some of the available agents have significant issues such as off-target toxicity and low efficiency of PTC suppression. Therefore, there is a need for therapeutic agents and methods for treatment of disorders associated with nonsense mutations.

Technology Overview

We have developed a platform of engineered tRNAs, termed ACE-tRNAs for AntiCodon Edited tRNAs. This technology recognizes the PTCs and reverts them to an amino acid during the translation of a mRNA resulting in functional gene products. ACE-tRNAs are encoded by small cDNAs and can be delivered by nanoparticles, DNA-protein complexes and by electroporation into various cell and tissue types in the body. ACE-tRNA cDNAs have shown efficient suppression of PTC encoded in different human cell lines.

Benefits

The ACE-tRNAs exhibit high efficiency of entry into cells, persistent expression of the ACE-tRNAs inside the cells, and high efficiency of PTC suppression (readthrough) with minimal suppression of natural termination codon, indicating exciting therapeutic promise with low off-target toxicity. Large quantities can be made easily in vitro so they are not contaminated with endotoxins like traditional cDNA. Finally, they are resistant to cellular exonucleases so they have extremely long half-lives in cells.

Applications

Cystic fibrosis, Duchenne muscular dystrophy, spinal muscular atrophy, infantile neuronal ceroid lipofuscinosis, β-thalessemia, cystinosis, X-linked nephrogenic diabetes insipidus, Hurler syndrome, Usher syndrome, and polycystic kidney disease.