Construction of a cell model of collagen metabolism abnormality induced by knockout of sparc gene and application thereof

By designing and validating sgRNA sequences and recombinant vectors targeting the human SPARC gene, the problem of low sgRNA editing efficiency in HeLa cells using CRISPR-Cas9 technology was solved, and an efficient SPARC gene knockout model was constructed to study the function of SPARC in tumor cell migration and collagen metabolism.

CN122344574APending Publication Date: 2026-07-07GUANGXI UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
GUANGXI UNIV
Filing Date
2026-04-14
Publication Date
2026-07-07

AI Technical Summary

Technical Problem

Existing CRISPR-Cas9 technology suffers from problems such as low sgRNA editing efficiency, strong design blindness, low screening efficiency, and lack of efficient and specific sgRNA tools when constructing SPARC gene knockout models, especially in human cervical cancer HeLa cells where efficient knockout is difficult to achieve.

Method used

A sgRNA sequence (sgRNA1, sgRNA2, sgRNA3) targeting the human SPARC gene was provided. Combined with the recombinant expression vector PX459 V2.0, a homozygous SPARC gene knockout model was constructed by transfecting HeLa cells, and the knockout effect was verified by PCR and T7EI restriction enzyme digestion.

Benefits of technology

Efficient and specific SPARC gene knockout was achieved in HeLa cells, significantly shortening the research cycle and providing stable experimental materials for in-depth research on the function of SPARC in tumor cell migration and collagen metabolism.

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Abstract

The present application relates to the technical field of biotechnology, and provides a CRISPR-Cas9 system sgRNA sequence capable of efficiently targeting and knocking out a human SPARC gene, a recombinant vector containing the sgRNA sequence, and a construction method and application demonstration of a SPARC gene knockout cell model.The sgRNA sequence can mediate a high-specificity and high-efficiency SPARC gene knockout effect, effectively solving the problems of strong blindness, low screening efficiency and lack of reliable tool sequences in the prior art sgRNA design for the gene.The SPARC gene knockout Hela cell line constructed by using the sgRNA sequence and the recombinant vector has important technical value and practical significance for studying the extracellular matrix network regulation and cell migration ability.
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