Lentiviral Vector-Mediated In Vitro Down-Regulation of Porcine Somatostatin Expression Using shRNA

Abstract:

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Two shRNA sequences against porcine somatostatin (SST) were designed using software available on the NCBI website. The designed RNA sequences were chemically synthesized and cloned into lentiviral vectors (LV-siRNA1 and LV-siRNA2). Porcine somatostatin cDNA was amplified and cloned into pcDNA3.1 (pcDNA3.1-SST). LV-siRNA1 or LV-siRNA2 was cotransfected with pcDNA3.1-SST into NIH3T3 cells. Real-time RT-PCR for the detection of SST mRNA, revealed that LV-siRNA1 and LV-siRNA2 suppressed SST expression by 87.9% and 86.3% (P < 0.01), respectively. In addition, radioimmunoassay (RIA) for direct detection of SST indicated that the suppression ratios for LV-siRNA1 and LV-siRNA2 were 55.1% and 51.6% (P < 0.01), respectively. These data showed that the 2 shRNA sequences were effective in suppressing SST expression and may provide an approach to down-regulate both in vitro and in vivo expression of porcine SST.

Info:

Periodical:

Advanced Materials Research (Volumes 343-344)

Edited by:

David Wang

Pages:

1248-1254

DOI:

10.4028/www.scientific.net/AMR.343-344.1248

Citation:

J. H. Ding et al., "Lentiviral Vector-Mediated In Vitro Down-Regulation of Porcine Somatostatin Expression Using shRNA", Advanced Materials Research, Vols. 343-344, pp. 1248-1254, 2012

Online since:

September 2011

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Price:

$35.00

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