Abstract: 【Objective】 Playing an important role in fruit ripening and softening, In order to inactivate the tomato cell wall glycoside hydrolase gene β-Hex by mutation of CRISPR-Cas system and inhibit the degradation of n-glycoprotein and the production of free n-glycan, to reduce the degree of hydrolytic softening of cell wall and regulate the excessive softening of tomato fruit lay the foundation for further research.CRISPR-Cas gene editing system will be used to regulate tomato fruit softening by site-specific mutation. 【Methods】 The characteristics, conserved domain, genome structure, digital expression profile and promoter cis-acting elements of Slβ-Hex polypeptide were systematically analyzed using biological online tools, and suitable sgRNAs were designed and screened according to CRISPR-Cas9 target design principles to promote the construction of tomato gene editing efficient expression vector. 【Results】 Tomato SLβhex gene was located on the complementary chain of tomato chromosome 1, and composed of 2 exons and 1 intron, encoding 575 amino acids. Transcriptome analysis of Slβ-Hex based on RNA-seq showed that Slβ-Hex was mainly expressed in the fruit of tomato, especially in the peel. The positive chain of the promoter distributed 7 Sgrnas, negative chain distributed 20 Sgrnas, of which 10 Sgrnas contained 16 cis-acting elements. 【Conclusion】 High-specific sgRNA is selected for gene editing to mutate the cis-element sequence and inhibit the expression of Slβ-Hex.