US20250325698
2025-10-23
Human necessities
A61K48/005
The patent application describes therapeutic methods and compositions using CRISPR Type V systems, specifically focusing on RNA guides that include both ribonucleotide and deoxyribonucleotide bases. These systems are applied in genome editing of various cell types, including somatic cells, induced pluripotency stem cells (iPSCs), and germline or embryonic cells. The aim is to facilitate xenotransplantation for organ and tissue therapies.
The CRISPR-Cas12 system, a part of the Type V CRISPR systems, is utilized for precise genome editing. This involves deleting, inserting, mutating, or substituting specific nucleic acid sequences. The Cas12a protein, which lacks the HNH nuclease domain found in Cas9, is central to this process. It can introduce double-strand breaks (DSBs) in DNA, which are then repaired through various mechanisms such as non-homologous end joining or homology-directed repair.
The application outlines methods for treating diseases characterized by aberrant gene expression. This involves introducing a nucleoprotein complex containing Cas12a and a CRISPR guide molecule into somatic cells. The process utilizes lipid nanoparticles to deliver these complexes, facilitating the insertion of donor polynucleotides that correct gene expression. These therapeutic strategies are applicable both ex vivo and through systemic administration.
The disclosed compositions include a nucleoprotein complex and a donor polynucleotide encapsulated in lipid nanoparticles. These nanoparticles may contain various lipids, such as DSPC, DPPC, and others, to ensure effective delivery. The compositions are designed to target specific genes associated with diseases, enabling precise genome modifications that can restore normal gene function.
The application also explores the use of genetically modified iPSCs for disease treatment. The process involves editing iPSCs with CRISPR-Cas12, followed by differentiation into cell types affected by the disease. These differentiated cells are then administered to patients. The reprogramming of iPSCs is achieved through various methods, including the introduction of specific mRNAs and exposure to different inhibitors and compounds to enhance differentiation into target cell types.