Functional consequences of knocking down porcine prostaglandin synthases in SK-6 swine kidney cell line.

We hypothesized that in vitro knock-down of the previously cloned genes of prostaglandin synthases will result in a reduction of synthesis, and thus secretion of their respective products, i.e., prostaglandin (PG) E2 or PGF2α. For this purpose, we designed short hairpin RNA (shRNA)-encoding constructs to knock down porcine mPGES-1 and PGFS (also named as AKR1CL1 in GenBank) and used them to transfect swine kidney SK-6 cells. Knocking down PGFS or mPGES-1 transcripts resulted in at least 50% inhibition of protein expression of each respective enzyme, as well as a reduction in the production of their respective prostaglandin, PGF(2α) or PGE(2). These results confirmed the identities of PGFS and mPGES-1. Moreover, they illustrate a unique opportunity to use the gene knock-down constructs in primary endometrial cells in order to study their biological roles in the porcine endometrium, particularly during the establishment of pregnancy.

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