Gene expression profile at single cell level of islets from two ß-cell specific knockout mouse models (ßZKO-Mip and ßZKO-Rip), littermate controls (Zzef1f/f), and the MIP-Cre mice fed with normal-chow diet (NCD).
Islet consists of multiple cell types that work in concert to orchestrate their endocrine function. To rule out of direct effect of diet or phenotype, we performed comparative single-cell RNA-sequencing (scRNA-seq) on 12-week ßZKO-Mip, ßZKO-Rip mice, littermate controls (Zzef1f/f) and MIP-Cre mice islets fed with normal-chow diet (NCD) by ?Chromium Next GEM Single Cell 3' Reagent Kits. Overall design: Briefly, two ß-cell specific knockout mouse models (ßZKO-Mip and ßZKO-Rip) were generated by crossing the C57/BL6N mice harboring floxed exon 15 (Zzef1f/f) with MIP-Cre or RIP-Cre mice. Each islet sample was gathered from no less than three mice with the same genotype. Then, purified islets were treated with 2 ml 0.05% TryLE at 37? for 4 min, followed by gently pipetting for about 20 times. The cell suspension is stained with 0.4% trypan blue to assess cell viability under microscopic observation. The sorted cells with greater than 85% viability are qualified for 3' digital gene expression by profiling ~10,000 individual cells per sample using 10x Chromium Single-Cell 3' and V(D)J library construction (10x Genomics, USA), according to the manufacturer's instructions.