Comparison of Ildr2 knockdown by (Adv-shRNA) and Ildr2 knockout (Adv-Cre) in mouse liver

We observed that Ildr2 knockdown via adenovirally-delivered shRNA (Adv-Ildr2 shRNA) causes hepatic steatosis in mice. Acute, liver-specific Ildr2 knockout mice generated by infecting C57BL/6J mice segregating for a floxed allele of Ildr2 with adenoviral Cre recombinase (Adv-Cre) do not develop hepatic steatosis. However, administration of Adv-Ildr2 shRNA to Ildr2 knockout mice (Ildr2floxed; albumin-cre) did cause hepatic steatosis, indicating that the Adv-Ildr2 shRNA had apparent “off-target” effects on gene(s) other than Ildr2. To determine additional targets of Ildr2 shRNA that may regulate hepatic lipid metabolism, we performed RNA sequencing on liver samples from (1) Adv-Ildr2 shRNA knockdown mice, (2) Adv-lacZ shRNA control mice, and (3) Adv-Cre Ildr2 knockout mice. For the purposes of our study, we sought to identify genes downregulated by Adv-Ildr2 shRNA, but unchanged in Adv-lacZ shRNA control or Adv-Cre Ildr2 knockout mice. The hepatic steatosis observed in Adv-Ildr2 shRNA knockdown mice was likely due to shRNA targeting of other “off-target” genes. We propose that the genes candidates identified in this sequencing experiment may lead to identification of novel regulators of hepatic lipid metabolism. Liver tissue from male, BL6 mice, 10 wks old. Adv-lacZ shRNA (n=4), Adv-Ildr2 shRNA (n=4), Adv-cre (n=4) Submitter indicates the raw data files are no longer available.