2results about How to "High knockout efficiency" patented technology

The application discloses a double-sgRNA site knockout cassette suitable for fusarium oxysporum, a CRISPR / Cas9 system and a construction method and application thereof, and is based on a filamentous fungus gene knockout plasmid pFC332 to construct a new vector pFRCas9-G418 which is more suitable for gene knockout of the fusarium oxysporum. The application provides a method for knocking out genes by using the double-sgRNA site CRISPR / Cas9 system suitable for the fusarium oxysporum, the method can realize rapid knockout of genes of the fusarium oxysporum, the knockout period is shortened to 20 days, and the knockout efficiency is as high as 77.8%. The application successfully verifies the influence of the FCC1 gene knockout on the morphology of the fusarium oxysporum, the center of the strain presents a yellow solid, and the growth morphology of the strain is small, and the FCC1 gene can be used as a phenotype reporter gene of the fusarium oxysporum in the future, so as to verify gene editing.

The present application relates to an application of an inducible promoter in Clostridium beijerinckii gene editing, and belongs to the technical field of biotechnology and genetic engineering. The rhamnose inducible promoter from Clostridium beijerinckii is applied to the CRISPR / Cas9 gene editing system of Clostridium beijerinckii, and the gene knockout efficiency is significantly improved, and an efficient and controllable promoter tool is provided for genetic modification of Clostridium beijerinckii. The present application provides a solution for improving the gene editing technology of solventogenic Clostridium, and significantly improves the efficiency and feasibility of genetic operation in the industrial strain.