A novel technique for the synthesis of RNA polynucleotides incorporating modified uridine nucleotide analogs.

The development of RNA-based therapies and vaccines is currently facing critical challenges that include enhancing their stability and efficacy, addressing the non-specific incorporation of modified nucleotides, and reducing the potential immunogenic responses associated with these RNA therapies.

This technology outlines a method for synthesizing RNA polynucleotides with specific substitutions of uridine nucleotides with modified uridine nucleotide analogs, utilizing a DNA-dependent RNA polymerase lacking an aromatic amino acid at a critical position. It includes the use of modified uridine triphosphates like pseudouridine triphosphate, facilitating the generation of RNA polynucleotides for various applications including mRNA vaccines with enhanced safety profiles, therapeutic RNA molecules for gene therapy, and RNA-based diagnostic tools.

Optimization & Testing

• Allows for precise incorporation of modified nucleotides into RNA sequences.
• Utilizes a specifically engineered RNA polymerase for enhanced synthesis efficiency.
• Enables the production of RNA with reduced immunogenicity, beneficial for therapeutic applications.
• Offers versatility in the selection of modified uridine analogs.

Fleming AM, Burrows CJ. Nanopore sequencing for N1-methylpseudouridine in RNA reveals sequence-dependent discrimination of the modified nucleotide triphosphate during transcription. Nucleic Acids Res. 2023;51(4):1914-1926. doi:10.1093/nar/gkad044

Aaron Duffy
(801) 585-1377
aaron.duffy@utah.edu