Construction of SALL4 shRNA vectors and THP-1 cell transfection

VernacularTitle:婆罗双树样基因4 shRNA干扰载体的构建及其对THP-1细胞的转染
Author: Jiang WU
Publication Type:Journal Article
Keywords: Oncogenes; SALL4; shRNA; THP-1 cells; RNA,small interfering
From: Journal of Leukemia & Lymphoma 2010;19(1):12-15
CountryChina
Language:Chinese
Abstract: Objective To construct an efficient SALL4 shRNA vector and transfect it into THP-1 cells for investigating the effect of SALL4 in leukemia. Methods Four SALL4-specific siRNA to aim at different SALL4 mRNA target sites and a negative control siRNA were designed, and pGPU6/GFP/Neo/SALL4 shRNA vectors were constructed. THP-1 cells were transfected and the expression of SALL4 in shRNA detected, and blank control and negative control were also designed. Results The results of real time quantitive PCR and Western blotting both exhibited that the interference effect of pGPU6/GFP/Neo/SALL4 shRNA-B vector was optimal targeting to mRNA-1122 target site and down-regulated the expression of SALL4 more significant(P <0.05). Conclusion Successfully construction of SALL4 siRNA vector by choosing SALL4 shRNA-B would be useful to accomplish study of SALL4.