Abstract: A method for performing gene editing on a target site in a cell, specifically a method for performing gene editing on a target site of a cell genome, comprising: (a) providing a cell to be genetically edited; (b) introducing into the cell (i) a gene editing enzyme or the encoding nucleic acid thereof or a first expression vector expressing the gene editing enzyme; and (ii) gRNA or a second expression vector expressing the gRNA, and performing gene editing on a target site of the cell genome, the gRNA directing the gene editing enzyme to perform fixed-point cutting on the target site; the targeting sequence of the gRNA targeting comprises one or more of the sequences shown in SEQ ID. No. 1-9. The method enables efficient gene editing to be performed on the target site.

Abstract: Disclosed is an sgRNA guiding a PD1 gene for cleavage to achieve the efficient integration of exogenous sequences. The method for gene editing a PD1 gene in cells includes the steps of introducing a nuclease and an sgRNA into cells, and gene-editing the PD1 gene. The sgRNA guides the nuclease to cleave the PD1 gene and forms a broken site, at which an exogenous donor repair template can also be introduced, so that CAR-T elements can be directionally inserted at the specific site of the PD1 locus to construct enhanced CD19-CART cells with PD1 knockout in one step.

Abstract: Disclosed by the present invention are a thienocyclic compound represented by formula (I), a pharmaceutically acceptable salt or a hydrate thereof, a composition containing the thienocyclic compound, and a preparation method therefor.