Application of RNA interference on gene expression of LIMK2

Objective: To silence the expression of LIMK2 of Balb/c mouse primary osteoblasts by RNAi and the effect was assayed through RT-PCR and Western-blot. Methods: Balb/c mouse primary osteoblasts were transfected by three different LIMK2 specific siRNAs (named R1, R2 and R3) and a negative control siRNA, respectively, which were all chemically-synthesized. After transfected by 24h and 48h respectively, RT-PCR and Western-blot was performed to detect the LIMK2 expression level and the most efficient siRNA sequence was determined. Results: Compared with the negative control siRNA, all the three LIMK2 specific siRNAs could silence efficiently the expression of LIMK2 of Balb/c mouse primary osteoblasts. The highest silence efficiency was R3 that was more than 70% using chemical synthesized siRNA. Conclusion: The expression of LIMK2 gene in Balb/c mouse primary osteoblasts was successfully silenced by the siRNA numbered R3.