CRISPR-CAS9 METHOD FOR SPECIFIC KNOCKOUT OF SWINE PDX1 GENE AND SGRNA FOR USE IN TARGETING SPECIFICALLY PDX1 GENE

摘要

A method utilizing CRISPR-Cas9 for specific knockout of swine PDX1 gene and an sgRNA for use in targeting specifically PDX1 gene. The target sequence on the PDX1 gene of the sgRNA targeting specifically the PDX1 gene complies with the sequence permutation rule of 5'-N(20)NFF-3', where N(20) represents 20 consecutive nucleobases, wherein each N represents either A or T or C or G; the target sequence on the PDX1 gene is located at either the first exon coding region at the N-terminus of the PDX1 gene or a junction with an adjacent intron; and the target sequence on the PDX1 gene is unique. The sgRNA is for use in the CRISPR-Cas9 method for specific knockout of swine PDX1 gene, allows rapid, precise, highly efficient, and specific knockout of swine PDX1 gene, effectively solves the problem of extended cycle and high costs for constructing PDX1 gene-knockout swine.