Targeted delivery of CRISPR interference system against Fabp4 to white adipocytes ameliorates obesity, inflammation, hepatic steatosis, and insulin resistance

Validation of (dCas9/sgFabp4) + ATS-9R and (dCas9-KRAB/sgFabp4) + ATS-9R oligoplexes in adipocytes. (A) dCas9/sgNeg, dCas9/sgFabp4 #1,#4, dCas9-KRAB/sgFabp4 #1,#4 were complexed with ATS-9R in pre-adipocytes and mature adipocytes and transfected under optimized conditions. The Fabp4 mRNA level was normalized and calculated as a value relative to the nontreated cells. (B) Mature adipocytes were treated as mentioned above, and the lipid droplets were assessed by Oil Red O staining to evaluate the antilipogenesis effect through CRISPRi-mediated delivery of dCas9 and sgFabp4. (C) Agarose gel image showing the CRISPR interference repression activity measured by T7E1 assay after transfection of dCas9/sgFabp4, (dCas9/sgNegative) + ATS-9R, (dCas9/sgFabp4) + ATS-9R, and (dCas9-KRAB/sgFabp4) + ATS-9R in mature adipocytes. (M) Molecular weight size marker.