(1) Effects of RNAi in vitro: We found that RNAi treatments temporarily inhibited the expression of chLEPR mRNA and the protein in COS-7 cells. In CHO-chLEPR cell strains, the luciferase activity of RNAi treated group was significantly lower than that of control at 3, but not 24 hr after leptin loading (125 ng/ml). It seemed that the designed RNAi was effective to inhibition of the signaling transmission in the leptin receptors.
(2) Effects of RNAi in vivo: The designed RNAi did not affect the expression of chLEPR mRNA in diencephalon of chicks at 15 or 24 hr after injection. Similar to the expression, the injection did not influence on both feed intake and body weight gain in chicks.
(3) Effect of anti-sense oligonucleotide for leptin or NPY Y1 receptor (in vivo) : Similar to RNAi, anti-sense oligonucleotide for leptin receptor did not affect the expression of chLEPR mRNA, and feed intake and body weight gain in chicks. On the other hand, anti-sense oligonucleotide for NPY Y1 receptor suppressed the expression of the receptor mRNA at 15 hr after injection.
