RNA-Seq experiment of wild-type mouse livers with mLMRs knockdown and hLMRs rescue

Rescue experiment in wild-type mice offers a valuable mean to verify the functional similarity of lncRNAs. By suppressing mLMRs in mice and observing the impact on gene regulation, followed by overexpression of hLMRs, we can determine the ability of hLMRs to rescue the loss-of-function effects of mLMRs. Should hLMRs exhibit a reversal of the loss-of-function on transcriptome regulations, this would suggest that they have similar functions to mLMRs. To do this, we used adenovirus-delivered shRNAs to suppress the expression of mLMRs in the livers of wild-type mice, while overexpressing corresponding hLMRs as a rescue group. Wild-type mice (C57BL6, 12 weeks age) were injected with three groups of viruses, including control viruses for knockdown and overexpression (Lac+pAdv5, control group), mLMR knockdown viruses and control viruses for overexpression (mouse-lncRNA KD+pAdv5, mLMR knockdown group), and mLMR knockdown viruses and hLMR overexpression group (mouse-lncRNA KD+human-lncRNA OE, hLMR rescue group). The adenoviruses were intravenously delivered to mice at a dose of 5 × 10^8 PFU/mouse for overexpression virus or 1 × 10^9 PFU/mouse for knockdown virus. After a 5-hour fast (~9:00 am to 2:00 pm), the livers were harvested for RNA-Seq analysis after 7 days.