Construction of SALL4 shRNA vectors and THP-1 cell transfection
- Author:
- No author available
- Journal Title:
- JOURNAL OF LEUKEMIA & LYMPHOMA
- Issue:
- 1
- DOI:
- 10.3760/cma.j.issn.1009-9921.2010.01.004
- Key Word:
- 癌基因;SALL4;短发夹RNA;THP-1细胞;RNA,小分子干扰;Oncogenes;SALL4;shRNA;THP-1 cells;RNA,small interfering
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.
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