Evaluation the suppression of HMGB1 gene expression in Wharton jelly cells by CRISPR/CAS9

Abstract

Introduction: Epidermolysis bollusa is a heterogeneous skin disease that affects 500000 people around the world .yet there is no certain treatment for it but , new treatment strategies which are based on protein , fibroblast and bone marrow transplant , predict a good future . recently for the treatment of skin disease , the use of epidermal cells has been considered . HMGB1 is a protein that produced and realesed by immune cells such as macrophage , monocytes and dendritic cells . this marker is abundant in the epidermolysis bollusa blisters and it is also increased in other autoimmune disease. The aim of this study was to evaluate the suppression of HMGB1 (as a well-known marker in incidence and severity of epidermolysis bollusa) in invitro by CRISPR/CAS9 system .

Methods and materials: The vector containing the HMGB1_CRISPR / CAS9 genes was designed and synthesized along with GGFP marker and lenticular viral genes. After confirmation of the vector, the HECK293 cells were used for packaging. Finally, the medium containing the virus was collected at 24, 48 and 72 hours at three times and was poured onto Wharton jelly cells. The PCR steps were performed after a week and the results were statistically analyzed using SPSS software.

Results: The statistical data from the case and control groups were analyzed by SPSS17 software. Based on the results of this study, the expression of the HMGB1 and the wharton jelly cells in the normal state (control group) was 0.02 and the case group (suppression) of the gene By CRISPR 0.005 , which decreased by 4 times. These cultivars were reported statistically significant with P <0.05.

Discution: Based on the results of this experimental study, suppression of HMGB1 gene expression through a CRISPR system was successful and can be further investigated in animal studies.