Generation and Characterization of Cytochrome P450 2J3/10 CRISPR/Cas9 Knockout Rat Model

Abstract

Cytochrome P450 2J2 (CYP2J2) enzyme attracts more attention because it not only metabolizes clinical drugs, but also mediates the biotransformation of important endogenous substances and the regulation of physiological function. Although CYP2J2 is very important, few animal models are available to study its function in vivo. In particular, CYP2J gene knockout (KO) rat model for drug metabolism and pharmacokinetics is not available. In this report, the CRISPR/Cas9 technology was used to delete rat CYP2J3/10, the orthologous genes of CYP2J2 in humans. The CYP2J3/10 KO rats were viable and fertile and showed no any off-target effect. Compared with wild-type (WT) rats, the mRNA and protein expression of CYP2J3/10 in liver, small intestine and heart of KO rats were completely absent. At the same time, CYP2J4 mRNA expression and protein expression were significantly decreased in these tissues. Further in vitro and in vivo metabolic studies of astemizole, a typical substrate of CYP2J, indicated that CYP2J was functionally inactive in KO rats. The heart function indexes of WT and KO rats were also measured and compared. The myocardial enzymes including creatine kinase-muscle brain type (CK-MB), creatine kinase (CK) and CK-MB/CK ratio of KO rats increased by nearly 140%, 80%, and 60%, respectively. In conclusion, this study successfully developed a new CYP2J3/10 KO rat model, which is a useful tool to study the function of CYP2J in drug metabolism and cardiovascular disease.

Evaluation of Herb–Drug Interaction Between Danshen and Rivaroxaban in Rat and Human Liver Microsomes

“…For example, CYP2J2 transgenic mice have a reduced rate of arrhythmia induction (Westphal et al, 2013). Our previous study also found that knockout of CYP2J3/ 10 in rats led to heart disease (Lu et al, 2020). In addition, CYP2J2*7 allele has been reported to be associated with stroke and CVD (Wang SY.…”

Differentially Expressed Genes and Molecular Susceptibility to Human Age-Related Diseases

“…First, we used the PubMed database [ 75 ] and characterized each of the 39 novel PAG-related DEGs of the tame and aggressive rats ( Table 2 ) by answering the question as to how under- or overexpression of their human orthologs can aggravate or alleviate ARDs [ 86 , 87 , 88 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 100 , 101 , 102 , 103 , 104 , 105 , 106 , 107 , 108 , 109 , 110 , 111 , 112 , 113 , 114 , 115 , 116 , 117 , 118 , 119 , 120 , 121 , 122 , 123 , 124 , 125 , 126 , 127 , 128 , 129 , 130 , 131 , 132 , 133 , 134 , 135 , 136 , ...…”

Site-directed deuteration of dronedarone preserves cytochrome P4502J2 activity and mitigates its cardiac adverse effects in canine arrhythmic hearts

“…Transgenic mice with cardiac-specific CYP2J2 overexpression and subsequent elevated intracardiac EET levels, exhibit enhanced cardiac I Ca,L , K ATP , transient outward K + ( I to) currents, and shortened cardiac action potential duration without incidences of arrhythmogenesis or sudden cardiac death 44 . In parallel, cardiac-specific CYP2J3/10 −/− transgenic rats (rat isoforms of human CYP2J2) exhibited 75% loss in metabolic activity and 66.6% reduction in in vivo probe substrate clearance compared to WT rats 45 . Additionally, creatine kinase-muscle brain type, serum creatine kinase and their ratio were considerably increased indicative of myocardial injury and cardiomyocyte damage 46 .…”