稳定敲低PLEKHQ1基因细胞株的建立

Establishment of stable RAW and THP-1 cell strain with PLEKHQ1-knock-down

目的:建立稳定敲低PLEKHQ1基因的RAW和THP-1细胞株。方法:在293TX细胞中进行病毒的包装,用获得的高滴度慢病毒感染RAW和THP-1细胞,通过免疫印迹和荧光显微镜,检测病毒感染后RAW和THP-1细胞PLEKHQ1蛋白表达的变化。结果:PLEKHQ1-lentiviral sh RNA-1质粒在转染后表现出明显的干扰作用,感染后能明显下调RAW和THP-1细胞的PLEKHQ1蛋白表达。结论:稳定敲低PLEKHQ1基因的RAW和THP-1细胞株的建立,可为进一步研究PLEKHQ1在相关疾病中的功能奠定基础。

Objective:To establish RAW and THP-1cell strain which PLEKHQ1gene was stably knocked down.Methods:The lentiviral plasmids was transfected into293TX cells to achieve package.And then the packaged lentivirus of high titer were used to infect RAW and THP-1cell.Western blot and Fluorescence microscope were applied to detect and analyze the change of expression of PLEKHQ1protein after RAW and THP-1cell was infected by lentivirus.Results:PLEKHQ1-lentiviral shRNA1showed significant interference effect after it was transfected,and can obviously reduced expression of PLEKHQ1in RAW and THP-1cells.Conclusion:The establishment of RAW and THP-1cell strain with PLEKHQ1knock-down can lay a foundation for further researching the function of PLEKHQ1in relevant disease.