The generation of Angptl3 knockout mice was performed essentially as described above, except that a mix of two gRNAs and no ssODNs were used for embryo injections. gRNAs with predicted cleavage sites flanking the entire Angptl3 gene (protospacers 5′-AATTACTAAGAGTGTGACTC-3′ and 5′-TAATGCCAATCCACGAGCAT-3′) were used to cleanly delete the gene (~9.1 kb). PCR amplification around the junction of the predicted cleavage sites was used to confirm the deletion (F: 5′-TGCAGCTATCCCAATGAATGAG-3′ and R: 5′-AGAGAAACGACACCCTTCAC-3′). Wild-type and homozygous knockout littermates of both sexes from several litters, ~8 weeks of age, were used for plasma lipid measurements using Infinity Triglycerides Reagent (Thermo Fisher Scientific) and Infinity Cholesterol Reagent (Thermo Fisher Scientific).
