Integrases and methods of use for efficient insertion of DNA sequences
Mutant Bxb1 integrases targeting rDNA pseudo-att sequences enhance integration efficiency and expand integration sites, addressing limitations of current recombination-based methods by enabling precise and efficient large DNA cargo insertion in mammalian cells.
Patent Information
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- KOMO BIOSCIENCES INC
- Filing Date
- 2025-12-19
- Publication Date
- 2026-06-25
AI Technical Summary
Current recombination-based methods for generating stable protein-expressing cell lines face challenges such as identifying safe harbor genomic locations, accurate sequence placement, fidelity in targeting, multi-insertion limitations, DNA payload size, efficiency, and risks of unintended mutations or DNA breaks, particularly in mammalian cell culture.
Employing mutant Bxb1 integrases that target pseudo-att sequences found in ribosomal DNA (rDNA) for precise and efficient integration of large DNA cargos, utilizing engineered nucleic acid molecules with modified pseudo-att and attP sequences to enhance integration efficiency and diversify integration sites.
The use of mutant Bxb1 integrases allows for improved integration efficiency and expanded integration sites, enabling multi-gene insertions and therapeutic protein expression with reduced off-target effects and mutation risks.
Smart Images
Figure US2025060733_25062026_PF_FP_ABST