Enzymatic Ligation Technology
Enzymatic ligation builds a full-length RNA from short, high-purity fragments joined by an enzyme. We apply this approach in two ways: enzymatic siRNA synthesis for GMP-grade siRNA from several kilograms up to 200 kg, and GenClip™ for long RNA from 100 to 500 mer.
Two Applications of Enzymatic Ligation
Both technologies start from short fragments that can be synthesized with high purity, then join them enzymatically — for siRNA at manufacturing scale, and for long RNA of 100 to 500 mer.
Enzymatic siRNA Synthesis
Our technology utilizes enzyme-mediated ligation of multiple RNA fragments to produce target siRNA compounds with higher yield and purity compared to conventional solid-phase synthesis. This method enables the production of GMP-grade products in quantities ranging from several kilograms up to 200 kg.
The siRNA is divided into four short fragments produced with AJIPHASE™, our liquid-phase synthesis technology. Because impurities are removed in the enzyme ligation process, the purification step for the synthetic fragments can be omitted. This service is offered by Ajinomoto Co., Inc.
Patent: WO2020171092 — Method for Producing Modified Oligonucleotide Including Complementary Sequence
Advantages of Enzymatic siRNA Synthesis
High Purity
The short length of the synthesized fragments allows for high purity production. In the enzyme ligation process, impurities (such as N±1 mer) can be effectively removed, and minimal residual complementary strand remains after ligation.
High Productivity
High-quality intermediates enhance purification efficiency compared to general production methods, with high reproducibility and productivity even in large-scale production.
Reduced Environmental Impact
High productivity reduces the amount of raw materials used, lowering the environmental impact throughout the oligonucleotide production supply chain.
GenClip™ Long RNA Synthesis
GenClip™ enables the synthesis of long RNA (100–500 mer) with high yield and high quality. It is also usable for the synthesis of sgRNA, pre-miRNA and mRNA, and due to its high quality, long RNA produced with GenClip™ can also be used as analytical standards.
The combination of solid-phase synthesis and enzymatic ligation enables higher yield than conventional methods, and GenClip™ is applicable to various chemical modifications such as 2′-F, 2′-MOE and LNA. GenClip™ is currently available for research use (Non-GMP); GMP manufacturing is under development.
Patent: WO2024048684 — Method for Producing Nucleic Acid Molecule
GenClip™ Results
Examples of our work — from long RNA and sgRNA to a full-length mRNA that is translated in a cell-free assay.
Let’s discuss enzymatic ligation for your program
siRNA at scale. Long RNA with high quality.