INCREASED NUCLEIC ACID-GUIDED CELL EDITING VIA LEXA-RAD51 FUSION PROTEIN

• Main Authors: GANDER MILES, SPINDLER EILEEN, TIAN TIAN
• Format: Patent
• Language: Chinese; English
• Published: 25.02.2022
• 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 disclosure provides compositions and methods to increase the percentage of edited yeast cells in a cell population when employing nucleic acid-guided editing, and automated multi-module instruments for performing these methods. Thus, there is provided in one embodiment an editing vector for nucleic acid-guided nuclease editing in yeast, and comprising a promoter driving the transcription of an editing cassette comprising a guide nucleic acid and a donor DNA sequence; a yeast origin of replication; a bacterial origin of replication; a promoter for driving transcription of a coding sequence for a nuclease; a promoter for driving transcription of a selection marker; one or more LexA DNA binding sites; and a promoter driving transcription of a LexA-linker-Rad51 fusion protein. 本公开内容提供了当采用核酸指导的编辑时增加细胞群体中编辑的酵母细胞百分比的组合物和方法，以及用于进行这些方法的自动化多模块仪器。因此，在一种实施方案中提供了一种编辑载体，该编辑载体用于在酵母中进行核酸指导的核酸酶编辑，该编辑载体包含：驱动包含指导核酸和供体DNA序列的编辑盒转录的启动子；酵母复制起点；细菌复制起点；驱动核酸酶编码序列转录的启动子；驱动选择标记转录的启动子；一个或更多个LexA DNA结合位点；和驱动LexA-接头-Rad51融合蛋白转录