To prepare crypt-enriched intestinal epithelial cell suspension, isolated crypts were digested with digestion buffer (0.4 mg/ml Dispase and 0.1 mg/ml DNase in HBSS) for 20 min at 37°C with agitation, filtered through 70- and 40-μm-mesh cell strainers, washed with HBSS and resuspended with Advanced DMEM-F12 containing 10 mM HEPES, 1X Glutamax, 1X B27 supplement, 50 ng/ml EGF and 10 mM Y-27632. Dead cells were removed by EasySep Dead Cell Removal (Annexin V) kit (STEMCELL technologies). scRNA-seq libraries were generated using the Chromium Single Cell 30 Reagent Kit v2 (10X Genomics). Cells were loaded onto the 10X Chromium Single Cell Platform (10X Genomics),  (Single Cell 3′ library and Gel Bead Kit v.2) as described in the manufacturer's protocol (10x User Guide, Revision B). On average, approximately 10,000 cells were loaded. Generation of gel beads in emulsion (GEMs), barcoding, GEM-RT clean-up, complementary DNA amplification and library construction were all performed as per the manufacturer's protocol. Individual sample quality was checked using a Bioanalyzer Tapestation (Agilent). Qubit was used for library quantification before pooling. The final library pool was sequenced on an Illumina NovaSeq6000 instrument using a S1 flowcell.Average cell recovery for Prkcif/fPrkcif/f (WT1) was 13018 cells with 18442 mean reads per cell and 2390 median genes per cell. Average cell recovery for Prkczf/f(WT2) was 12670 cells with 43035 mean reads per cell and 2848 median genes per cell. Average cell recovery for Prkcif/f(WT3) was 18194 cells with 9809 mean reads per cell and 882 median genes per cell. Average cell recovery for Prkczf/f;Villin-Cre(ZKO) was 13546 cells with 47238 mean reads per cell and 3110 median genes per cell. Average cell recovery for Prkcif/f;Villin-Cre(LKO) was 17991 cells with 8902 mean reads per cell and 769 median genes per cell. Average cell recovery for Prkcif/fPrkcif/f;Villin-Cre(DKO) was 20848 cells with 11700 mean reads per cell and 1974 median genes per cell