TY - JOUR T1 - Construction and characterization of CRISPR/Cas9 knockout rat model of carboxylesterase 2a gene JF - Molecular Pharmacology JO - Mol Pharmacol DO - 10.1124/molpharm.121.000357 SP - MOLPHARM-AR-2021-000357 AU - Jie Liu AU - Xuyang Shang AU - Shengbo Huang AU - Yuan Xu AU - Jian Lu AU - Yuanjin Zhang AU - Zongjun Liu AU - Xin Wang Y1 - 2021/01/01 UR - http://molpharm.aspetjournals.org/content/early/2021/09/09/molpharm.121.000357.abstract N2 - Carboxylesterase 2 (CES2), an important metabolic enzyme, plays a critical role in drug biotransformation and lipid metabolism. Although CES2 is very important, few animal models have been generated to study its properties and functions. Rat Ces2 is similar to human CES2A-CES3A-CES4A gene cluster, with highly similar gene structure, function and substrate. In this report, CRISPR/Cas9 technology was firstly used to knock out rat Ces2a, a main subtype of Ces2 mostly distributed in liver and intestine. This model showed the absence of CES2A protein expression in liver. Further pharmacokinetic studies of diltiazem, a typical substrate of CES2A, confirmed the loss of function of CES2A both in vivo and in vitro. At the same time, the expression of CES2C and CES2J protein in liver decreased significantly. The body and liver weight of Ces2a knockout rats also increased, but the food intake did not change. Moreover, the deficiency of Ces2a led to obesity, insulin resistance and liver fat accumulation, which are consistent with the symptoms of nonalcoholic fatty liver disease (NAFLD). Therefore, this rat model is not only a powerful tool to study drug metabolism mediated by CES2, but also a good disease model to study NAFLD. Significance Statement Human CES2 plays a key role in the first-pass hydrolysis metabolism of most oral prodrugs as well as lipid metabolism. In this study, CRISPR/Cas9 technology was used to knock out Ces2a gene in rats for the first time. This model can be used not only in the study of drug metabolism and pharmacokinetics, but also as a disease model of NAFLD and other metabolic disorder. ER -