Construction of small interfering RNA targeting mouse vascular endothelial growth factor receptor-2: its serum stability and gene silencing efficiency in vitro / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To construct a small interfering RNA (siRNA) targeting mouse vascular endothelial growth factor receptor-2 (VEGFR2) and study its serum stability and gene silencing efficiency in vitro.</p><p><b>METHODS</b>The synthesized siRNA targeting VEGFR2 (siVEGFR2) diluted in RNase-free water was mixed at a 11 ratio with fresh serum and incubated at 37 degrees celsius;. Gel electrophoresis was performed to determine the integrity of siVEGFR2 incubated for different time lengths. Oligofectamine 2000 was used to mediate siVEGFR2 transfection of MS1 cells, and semi-quantitative RT-PCR was used to evaluate VEGFR2 gene silencing effect induced by the siRNA.</p><p><b>RESULTS</b>The naked siRNA incubated in serum underwent gradual degradation with prolonged incubation time and became virtually undetectable after 24 h. Transfection of MS1 cells with siVEGFR2 significantly down-regulated the expression of VEGFR2 mRNA in comparison with the blank group and control siRNA transfection group (P<0.001), while no significant difference was found in VEGFR2 mRNA levels between the latter two groups (P=0.157).</p><p><b>CONCLUSION</b>The naked siRNA is unstable in serum and not suitable for direct administration in vivo. The designed siRNA can effectively silence the VEGFR2 gene expression in MSI cells in vitro.</p>