Characterization of a Novel CYP1A2 Knockout Rat Model Constructed by CRISPR/Cas9

Abstract

Cytochrome P450 1A2 (CYP1A2) as one of the most important CYP isoforms is involved in the biotransformation of many important endogenous and exogenous substances. CYP1A2 also plays an important role in the development of many diseases because it is involved in the biotransformation of precancerous substances and poisons. Although the generation of Cyp1a2 knockout (KO) mouse model has been reported, there are still no relevant rat models for the study of CYP1A2-mediated pharmacokinetics and diseases. In this report, CYP1A2 KO rat model was established successfully by CRISPR/Cas9 without any detectable off-target effect. Compared with wild-type rats, this model showed a loss of CYP1A2 protein expression in the liver. The results of pharmacokinetics in vivo and incubation in vitro of specific substrates of CYP1A2 confirmed the lack of function of CYP1A2 in KO rats. In further studies of potential compensatory effects, we found that CYP1A1 was significantly up-regulated, and CYP2E1, CYP3A2 and LXRβ were down-regulated in KO rats. In addition, CYP1A2 KO rats exhibited a significant increase in serum cholesterol and free testosterone, accompanied by mild liver damage and lipid deposition, suggesting that CYP1A2 deficiency affects lipid metabolism and liver function to a certain extent. In summary, we successfully constructed the CYP1A2 KO rat model, which provides a useful tool for studying the metabolic function and physiological function of CYP1A2.

Construction and characterization of a humanized SLCO1B1 rat model with its application in evaluating the uptake of different statins

“…The mRNA levels of major absorptive transporters, efflux transporters, and CYP enzymes were detected to evaluate the compensatory effects of SLCO1B1 knockin. The detection method and primers used were consistent with our previous reports 26 , 27 .…”

Expression of Lipid Metabolism Genes Is Correlated With Immune Microenvironment and Predicts Prognosis in Endometrial Carcinoma

“…Although CYP1A2 is expressed predominantly in hepatocytes, little is known about its expression in extrahepatic tissues. CYP1A2 knockout mice displayed large increases in blood cholesterol and free testosterone, followed by minor liver damage and fat deposition, as demonstrated by Sun et al [25] . GDPD5, which gene is located on chromosome 11q13.5, was discovered as a member of the glycerophosphodiesterase (GDE) family, which is essential for glycerol metabolism, in humans.…”

Genome sequencing of 2000 canids by the Dog10K consortium advances the understanding of demography, genome function and architecture

“…10 b). Here, both sampled Keeshonds were homozygous LOF, providing a spontaneous canine model to study the compensatory effects of this gene knockout [ 131 , 132 ]. While the role of CYP1A2 is well-established, the roles of other potential drug targets examined in this study remain to be elucidated and require cautionary comment.…”