Lna dicer1

To perform Dicer1, Drosha and Dgcr8 knockdown experiments, 10 pl of 10 μM LNA-Dicer1, LNA-Drosha or LNA-Dgcr8 (Exiqon) was injected into GV oocytes and the expressions were checked in MII oocytes or zygotes. 10 pl of 10 μM LNA-L1 and LNA-asL1 were injected into MII oocytes to check the knockdown efficiency and the in vitro developmental competency of IVF embryos. Endo-siRNA mimics used in the study are small, chemically modified double-stranded RNAs that mimic endo-siRNAs and enable siRNA functional analysis by up-regulation of siRNA activity. Endo-siRNA inhibitors are small, chemically modified single-stranded RNA molecules designed to specifically bind to and inhibit endo-siRNA molecules and enable siRNA functional analysis by down-regulation of endo-siRNA activity.9 L1-siRNAs mimics and inhibitors were designed and synthesized by Invitrogen and mixed by 1:1, respectively. 10 pl of 20 μM mimics mixture or inhibitors mixture was injected into MII oocytes to perform IVF and detect the embryonic development. The embryos were cultured in porcine zygote medium-3 at 39°C in 5% CO₂ in air. The cleavage and blastocyst rates were assessed at 48 and 156 h after activation.