Engineered 3'o-kinase variants
Engineered 3'O-kinases overcome the limitations of NQP availability and cost by enhancing the synthesis of nucleoside quadriphosphates, enabling efficient large-scale oligonucleotide production.
Patent Information
- Application Number
- PCT/US2025/034048
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- Priority Date
- 2024-06-18
- Filing Date
- 2025-06-17
- Publication Date
- 2025-12-26
AI Technical Summary
The commercial availability and cost of nucleoside tetraphosphates (NQPs) with a phosphate at the 3' position of the sugar are limiting factors for large-scale synthesis of oligonucleotides, particularly in template-independent synthesis applications, as they are not widely available and prohibitively expensive.
Engineered 3'O-kinase polypeptides, derived from a variant of the wild-type dephospho-CoA kinase (CoaE) from Geobacillus stearothermophilus, are developed to add a phosphate group to the 3' position of natural or modified nucleoside triphosphates, enhancing activity, thermostability, selectivity, and reducing byproduct formation.
The engineered 3'O-kinases enable the efficient production of nucleoside quadriphosphates (NQPs), addressing the limitations of existing synthesis methods by improving yield and reducing costs, thus supporting large-scale oligonucleotide synthesis.