March 2015 (No. 1) Volume 15
Functional consequences of knocking down porcine prostaglandin synthases in SK-6 swine kidney cell line
Agnieszka Waclawik a,*, Adolfo Rivero-Müller b,c, Nafis A. Rahman b, Adam J. Ziecik a
a Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland
b Department of Physiology, University of Turku, 20520 Turku, Finland
c Faculty of Natural Sciences and Technology, Åbo Akademi University, Turku, Finland
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, PGF2α or PGE2. 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 endome-trial cells in order to study their biological roles in the porcine endometrium, particularly during the establishment of pregnancy.
Reproductive Biology 2015, 15, 1: 42-47
* Corresponding author at: Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland; e-mail address: firstname.lastname@example.org (A. Waclawik).