GFP and mCherry gene expression at single cell level of all liver cell types from B6J mice
Diacylglycerol acyltransferase (DGAT)-2 catalyzes the final step of triglyceride (TG) synthesis. DGAT2 deletion in mice lowers liver TGs and DGAT2 overexpression might have the opposite effect. Mouse DGAT2 was overexpressed in C57Bl/6J mice using adeno-associated virus 8 (AAV8 and AAV-DJ). The tissue specificity of AAV8 and AAV-DJ was first evaluated.  To confirm that AAV8 and AAV-DJ only infected hepatocytes and not other cells in liver, we carried out single cell sequencing of liver cells isolated from mice infected with AAV8-mCherry and AAV-DJ-GFP. The mCherry and GFP signals were highly expressed in hepatocytes and not in stellate, endothelial, or Kupffer cells. Overall design: An 8-week-old C57BL/6J male mouse was injected retro-orbitally with an AAV-DJ virus that was engineered to express GFP and AAV-8 engineered to express mCherry. The mouse was fed a chow diet ad lib and sacrificed in 3 weeks after virus injection. Hepatocytes and non-parenchymal liver cells (NPCs) were isolated for the sc-RNA seq. New custom mice gene reference genomes were created by adding both GFP and mCherry into the existing mouse gene reference genomes. GFP and mCherry gene expression were assessed in both hepatocytes and NPCs.