Development and characterization of MDR1 (MDR1a/B) CRISPR/Cas9 knockout rat model

Chenmeizi Liang, Junfang Zhao, Jian Lu, Yuanjin Zhang, Xinrun Ma, Xuyang Shang, Yongmei Li, Xueyun Ma, Mingyao Liu, Xin Wang

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated protein-9 nuclease (Cas9) technology is widely used as a tool for gene editing in rat genome site-specific engineering. Multidrug resistance 1 [MDR1 (also known as P-glycoprotein)] is a key efflux transporter that plays an important role not only in the transport of endogenous and exogenous substances, but also in tumor MDR. In this report, a novel MDR1 (Mdr1a/b) double-knockout (KO) rat model was generated by the CRISPR/Cas9 system without any off-target effect detected. Western blot results showed that MDR1 was completely absent in the liver, small intestine, brain, and kidney of KO rats. Further pharmacokinetic studies of digoxin, a typical substrate of MDR1, confirmed the deficiency of MDR1 in vivo. To determine the possible compensatory mechanism of Mdr1a/b (2/2) rats, the mRNA levels of the CYP3A subfamily and transporter-related genes were compared in the brain, liver, kidney, and small intestine of KO and wild-type rats. In general, a new Mdr1a/b (2/2) rat model has been successfully generated and characterized. This rat model is a useful tool for studying the function of MDR1 in drug absorption, tumor MDR, and drug target validation.

Original languageEnglish
Pages (from-to)71-79
Number of pages9
JournalDrug Metabolism and Disposition
Volume47
Issue number2
DOIs
StatePublished - Feb 2019

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