New strategies for precision genome editing

• Main Authors: JONGEDIJK ERIK, VAGHCHHIPAWALA ZARIR, NIESSEN MARKUS, HUMMEL AARON, WELTMEIER FRIDTJOF, GLENZ RENE
• Format: Patent
• Language: Chinese; English
• Published: 14.08.2020
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The present invention relates to improved methods for precision genome editing (GE), preferably in eukaryotic cells, and particularly to methods for GE in cells with specifically altered expression ofPolymerase theta and altered characteristics of at least one further enzyme involved in a non-homologous end-joining (NHEJ) DNA repair pathway. Further provided are cellular systems and tools relatedto the methods provided. Specifically, methods are provided, wherein Polymerase theta and NHEJ blockage and/or GE are performed in a transient way, so that the endogenous Polymerase theta and cellular NHEJ machinery is easily reactivated after a targeted edit, and/or without permanent integration of certain editing tools. 本发明涉及优选在真核细胞中用于精确基因组编辑(GE)的改进方法，特别涉及用于通过特别地改变的聚合酶θ表达以及参与非同源末端连接(NHEJ)DNA修复途径的至少一种其他酶的特性，在细胞中GE的方法。还提供了与所提供的方法有关的细胞系统和工具。具体地，提供了方法，其中聚合酶θ和NHEJ阻断和/或GE以瞬时方式进行，使得内源聚合酶θ和细胞NHEJ机制在靶向编辑之后容易重新活化，和/或不需要某些编辑工具的固定整合。