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Cytochrome P450 (CYP) 3A accounts for nearly 30% of the total CYP enzymes in the human liver and participates in the metabolism of over 50% of clinical drugs. Moreover, CYP3A plays an important role in chemical metabolism, toxicity, and carcinogenicity. New animal models are needed to investigate CYP3A functions, especially for drug metabolism. In this report,  Cyp3a1/2  double knockout (KO) rats were generated by CRISPR-Cas9 technology, and then were characterized for viability and physiological status. The  Cyp3a1/2  double KO rats were viable and fertile, and had no obvious physiological abnormities. Compared with the wild-type (WT) rat,  Cyp3a1/2  expression was completely absent in the liver of the KO rat.  In vitro  and  in vivo  metabolic studies of the CYP3A1/2 substrates indicated that CYP3A1/2 was functionally inactive in double KO rats. The  Cyp3a1/2  double KO rat model was successfully generated and characterized. The  Cyp3a1/2  KO rats are a novel rodent animal model that will be a powerful tool for the study of the physiological and pharmacological roles of CYP3A, especially in drug and chemical metabolism  in vivo .

CRISPR knockout rat cytochrome P450 3A1/2 model for advancing drug metabolism and pharmacokinetics research