An experimental study on silencing the expression of LIMK2 gene by small interfering RNAs in mouse primary osteoblasts

doi:10.3877/cma.j.issn.1674-1366.2010.02.005

Abstract

Abstract:

Objective

To silence the expression of LIMK2 gene in primary mouse osteoblasts by small interfering RNAs （siRNAs） and evaluate their inhibition efficiency.

Methods

Three siRNAs which target mouse LIMK2 gene were chemically synthesized and named R1， R2 and R3， respectively. The siRNAs were transfected into primary osteoblasts by liposome. Reverse transcriptase polymerase chain reaction （RT-PCR） and Western blot analysis were performed to test the LIMK2 gene expression and then the most efficient siRNA sequence was chosen.

Results

The mRNA and protein levels of LIMK2 in the groups using R3 siRNA were much lower than those of other groups. Compared with the negative control group， the relative expression of LIMK2 mRNA in the the groups using R3 siRNA were 19.30% （24 h after transfection） and 18.63%（48h after transfection）， respectively. Compared with the blank control group， the relative expression of LIMK2 protein in the Group 7 and Group 8 using R3 siRNA were only 1.32%（24 h after transfection）and 1.19% （48 h after transfection）， respectively.

Conclusion

The expression of LIMK2 gene in mouse primary osteoblasts can be successfully silenced by the R3 siRNA.

Key words: LIMK2 gene, RNA interference, Osteoblasts

You-rui LI, Feng QIN, Yi-ping ZHANG, Minfeng SHAO, Rui CHEN, Yun SHEN, Qiang FU. An experimental study on silencing the expression of LIMK2 gene by small interfering RNAs in mouse primary osteoblasts[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2010, 4(02): 133-139.

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URL: https://zhkqyxyjzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-1366.2010.02.005

https://zhkqyxyjzz.cma-cmc.com.cn/EN/Y2010/V4/I02/133

Figures/Tables 8

References 12

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引物	序列	引物大小
LIMK2	Forward：5-TCTGCTGACCGAGTACATCG-3	284 000
	Reverse：5-TGCGTAAGGTGCGTTTCT-3
GAPDH	Forward：5-ACCACAGTCCATGCCATCAC-3	450 000
	Reverse：5-TCCACCACCCTGTTGCTGTA-3

引物	序列	引物大小
LIMK2	Forward：5-TCTGCTGACCGAGTACATCG-3	284 000
	Reverse：5-TGCGTAAGGTGCGTTTCT-3
GAPDH	Forward：5-ACCACAGTCCATGCCATCAC-3	450 000
	Reverse：5-TCCACCACCCTGTTGCTGTA-3

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