Engineered Acr proteins for modulating CRISPR activity

• Main Authors: RABUKA DAVID, CHAILLARD AURELIEN, DETARCA, LIVIUS, M., A, SCHEEL MICHAEL
• Format: Patent
• Language: Chinese; English
• Published: 27.05.2025
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

Compositions and methods are provided that include engineered anti-CRISPR (Acr) proteins that can achieve a balance in which the Cas protein retains activity to perform targeting functions at a target nucleic acid (e.g., DNA cleavage for gene editing applications), but is inhibited by the subject Acr protein to a degree to reduce off-target activity, such that the subject Acr protein may not inhibit off-target activity. Thus, the ratio of target to off-target nucleic acid targeting events is increased. In some cases, the subject compositions (e.g., subject systems) include a CRISPR nuclease or a nucleic acid encoding the same. Methods of using the subject Acr polypeptides with CRISPR nuclease enzymes to achieve target nucleic acid modification are provided. 提供了包括工程化抗CRISPR(Acr)蛋白的组合物和方法，所述工程化抗CRISPR(Acr)蛋白可以实现平衡，其中Cas蛋白保留活性以执行在靶核酸靶向功能(例如，用于基因编辑应用的DNA切割)，但是被主题Acr蛋白抑制到降低脱靶活性的程度，从而使得在靶与脱靶核酸靶向事件的比率增加。在一些情况下，主题组合物(例如，主题系统)包括CRISPR核酸酶或编码它的核酸。提供了将主题Acr多肽与CRISPR核酸酶一起使用以实现靶核酸修饰的方法。