Evaluation the suppression of VEGF gene expression inMCF7 cells by CRISPR/CAS9

Abstract

Introduction:

Making new blood vessels is one of the requirements for tumor growth and invasion. VEGF is one of the most important growth factors in angiogenesis, which plays an important role in the development of vascular and vasodilatation. Genomic engineering using CRISPER / Cas9 technology has dramatically improved the effectiveness and ease of targeted genetic modification. And has revolutionized in many aspects of biological and medical research in recent years The aim of this study was to control the expression of the VEGF gene using the CRISPR / cas9 system in breast cancer cells of MCF7. Cells were made from cellular storage of the new science university. After laboratory tests, VEGF gene expression was compared with the control group. Statistical data were analyzed with SPSS17 and P <0.05.

Methods and materials:

The vector containing the VEGF-_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 MCF7 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 VEGF and the MCF7 cells in the normal state (control group) was 27.3 and the case group (suppression) of the gene By CRISPR 0.65, which decreased by 5 times. These cultivars were reported statistically significant with P <0.05. (Figures 1 and 2) Discuss:

Based on the results of this experimental study, suppression of VEGF gene expression through a CRISPR system is a successful process and can be further investigated in animal studies.