Engineered split luciferases
Split luciferases with specific amino acid sequences and secondary structures enhance biosensor performance for detecting cellular events, addressing the need for improved molecular probes with high specificity and sensitivity.
WO2026090429A1PCT designated stage Publication Date: 2026-04-30MONOD BIO INC
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Patent Information
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- MONOD BIO INC
- Filing Date
- 2025-10-23
- Publication Date
- 2026-04-30
AI Technical Summary
Technical Problem
Existing luciferases are not sufficiently engineered for use as molecular probes with high specificity and sensitivity in detecting cellular events, requiring improved biosensors.
Method used
Development of split luciferases, comprising a larger and smaller component that form a complementing pair upon proximity, with specific amino acid sequences and secondary structures to enhance luciferase activity in complementation assays.
Benefits of technology
The split luciferases provide high specificity and sensitivity as biosensors, enabling effective detection of cellular events without the need for excitation, reducing background signals and phototoxicity.
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Abstract
The present disclosure provides polypeptides that can be used in a complementation assay as a biosensor. In certain aspects, the polypeptide is a larger component of a split luciferase, wherein the luciferase is derived from LuxSit. The polypeptide may comprise an amino acid sequence having at least 85% identity to the amino acid sequence: of SEQ ID NO:1 and comprises D at position 30, N at position 46, R at position 58, R at position 61, F at position 64, T at position 68, E at position 101, and A at position 103; G at position 9, D at position 30, D at position 54, F at position 64, K at position 68, and E at position 101; D at position 30, D at position 46, K at position 68, and E at position 101; S at position 9, D at position 30, L at position 43, D at position 46, G at position 54, F at position 64, K at position 68, E at position 101, and T at position 103; D at position 30, D at position 46, F at position 64, K at position 68, E at position 101, and T at position 103; N at position 30, D at position 46, E at position 101, and A at position 103; or D at position 30, N at position 46, F at position 64, T at position 68, E at position 101, and A at position 103, wherein the numbering of the amino acid positions is based on the numbering of the amino acids in SEQ ID NO:1.
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