RIBOZYME AND OMICS

PLATFORM TECHNOLOGY

RNA Replacement Enzyme Circular RNA

Circular RNA (CircRNA)

Advantages of circular RNA technology

Stability : non-exposure of 5’ or 3’ end for exonuclease activity

Safe : equivalent efficacy with smaller doses to reduce unwanted innate immunity

Efficient translation : Increased ribosome recycling & the protein expression due to the adjacent locations between start codon and stop codon

Application of the Circular RNA technology

mRNA therapeutics

mRNA vaccine (prophylaxis, anti-cancer)

Protein expressing mRNA for the CAR-based cell treatments

Anti-miRNA treatments

→ Replacing the linear RNA’s position

Advantages of Rznomics’ Circular RNA platform

Rznomics’ Method Chemical- & Ligase-Based
Methods		 PIE (Permuted intron exon)-Based
Methods
Ease of circRNA production 1 step ≥ 2 Step ≥ 1 Step
Yield after
purification		 Comparable or better compared to PIE method Not reported, but some issues
(Multimer produced by intermolecular interaction between RNAs and splint DNA)		 Not reported
Strategy to increase
the efficiency		 Target site screening
/
Optimization of P1/P10 & AS/ABS		 - Optimization of complementary sequences
Purification method
& purity		 Established IP-RP (Ion Pair-Reverse Phase) HPLC method Purification issue caused by splint DNA (Strong interaction with RNAs) SEC (Size Exclusion Chromatography) HPLC method
Extraneous
sequences/structures		 No No Yes

New Circular RNA Platform for RNA Therapeutics and Vaccines

Distinction compared to other in vitro circRNA preparation technologies
  • Circular RNA formation by Self-Targeting & Splicing (STS) Reaction (Any target sequences: 5’-NNNNNU-3’ in GOI)
  • Self-circularization can be completed during IVT
  • Avoids leaving unwanted sequences/structures (e.g. intronic scar) in the circRNA
  • Options for optimization of self-circularization efficiency (Target site screening, Optimization of P1/P10 & AS/ABS)

Rznomics’ Strategy for Circular RNA Preparation

Circularization by End-to-End Self-Targeting & Splicing (STS) Reaction
  • ASAntisense Sequence
  • ABSAntisense Binding Sequence
  • GOIGene of Interest