US20250320490
2025-10-16
Chemistry; metallurgy
C12N15/11
The patent application details innovative compositions and methods involving nucleic acid-targeting nucleic acids for genome engineering. These processes enable precise alterations in genetic sequences, which can include deletions, insertions, mutations, or substitutions. The focus is on both genomic and non-genomic nucleic acid engineering, utilizing nucleases to create specific sites for modification. This technology is significant for its potential applications in various fields, including genetics and molecular biology.
Genome engineering is a sophisticated process that involves altering specific nucleic acid sequences within a genome. This can be achieved by using nucleases to introduce double-stranded breaks at targeted sites, which are then repaired by cellular mechanisms such as non-homologous end joining (NHEJ). The application also considers the engineering of non-genomic nucleic acids, expanding the scope of potential modifications. These techniques have broad implications for research and therapeutic developments.
A key aspect of the disclosure is the engineering of nucleic acid-targeting nucleic acids, particularly through mutations in the P-domain. These engineered nucleic acids can include linker sequences that enhance their ability to hybridize with target sequences. The P-domain mutations can be configured to improve binding specificity and reduce non-specific interactions, thereby increasing the efficacy of genome modifications. The application also discusses the potential for these engineered nucleic acids to bind with greater specificity and lower dissociation constants than their unmodified counterparts.
The engineered nucleic acid-targeting nucleic acids can feature various configurations of the P-domain, which may include mutations, insertions, or deletions. These alterations are designed to optimize binding to different protospacer adjacent motifs, enhancing the precision of genome targeting. The application mentions specific motifs and configurations, indicating a high level of customization available for different genetic targets. Additionally, the engineered nucleic acids can form complexes with site-directed polypeptides, further extending their functionality.
The patent outlines methods for using these engineered nucleic acids to modify target nucleic acids. This can involve cleaving or altering the transcription of the target sequence, as well as inserting donor polynucleotides. Such methods hold promise for advancing genetic research and developing new therapeutic strategies. The ability to precisely edit genetic material opens up possibilities for treating genetic disorders, improving crop resistance, and more, highlighting the transformative potential of this technology.