Generation and Characterization of a CYP2C11-Null Rat Model by Using the CRISPR/Cas9 Method

Wei, Yuan; Yang, Li; Zhang, Xiaoyan; Sui, Danjuan; Wang, Chang-Suo; Wang, Kai; Shan, Mangting; Guo, Dayong; Wang, Hongyu

doi:10.1124/dmd.117.078444

Cited by 22 publications

(19 citation statements)

References 45 publications

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“…Using a CYP2C11 -null rat model that we had previously generated (Wei et al, 2018), here we examined how CYP2C11 influences EET function in vivo. We challenged CYP2C11 -null and wild-type (WT) Sprague-Daly (S-D) rats with a high-salt diet to understand how blood pressure would change in response.…”

Section: Introductionmentioning

confidence: 99%

CYP2C11 played a significant role in down-regulating rat blood pressure under the challenge of a high-salt diet

Liu¹,

Sui²,

Ye³

et al. 2019

PeerJ

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Background Arachidonic acid (AA) is oxidized by cytochrome P450s (CYPs) to form epoxyeicosatrienoic acids (EETs), compounds that modulate ion transport, gene expression, and vasorelaxation. Both CYP2Cs and CYP2Js are involved in kidney EET epoxidation. Methods In this study, we used a CYP2C11-null rat model to explore the in vivo effects of CYP2C11 on vasorelaxation. For 2 months, CYP2C11-null and wild-type (WT) Sprague-Dawley rats were either fed normal lab (0.3% (w/w) sodium chloride) or high-salt (8% (w/w) sodium chloride) diets. Subsequently, an invasive method was used to determine blood pressure. Next, western blots, quantitative PCR, and immunohistochemistry were used to determine renal expression of CYPs involved in AA metabolism. Results Among CYP2C11-null rats, a high-salt diet (females: 156.79 ± 15.89 mm Hg, males: 130.25 ± 16.76 mm Hg, n = 10) resulted in significantly higher blood pressure than a normal diet (females: 118.05 ± 8.43 mm Hg, P < 0.01; males: 115.15 ± 11.45 mm Hg, P < 0.05, n = 10). Compared with WT rats under the high-salt diet, western blots showed that CYP2C11-null rats had higher renal expression of CYP2J2 and CYP4A. This was consistent with the results of immunohistochemistry and the qPCR, respectively. The two rat strains did not differ in the renal expression of CYP2C23 or CYP2C24. Conclusion Our findings suggested that CYP2C11 plays an important role in lowering blood pressure under the challenge of a high-salt diet.

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Section: Introductionmentioning

confidence: 99%

CYP2C11 played a significant role in down-regulating rat blood pressure under the challenge of a high-salt diet

Liu¹,

Sui²,

Ye³

et al. 2019

PeerJ

Self Cite

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“…However, all models showed compensatory upregulation or downregulation of other P450s involved in drug metabolism. Moreover, these models also showed other differences, e.g., altered serum concentrations of testosterone (Lu et al, 2017) or alkaline phosphate (Wang et al, 2016) and reduced fertility (Wei et al, 2018). Some of these differences can be attributed to the function of the knocked out gene, e.g., CYP3A and testosterone metabolism, whereas others are unexpected.…”

Section: Downloaded Frommentioning

confidence: 98%

“…Unfortunately, no functional characterization was done and to our knowledge no follow-up studies with these in vivo models have been published. In other models Cyp2e1 (Wang et al, 2016) or Cyp2c11 (Wei et al, 2018) were knocked out in Sprague-Dawley rats. The CRISPR-Cas9 technology has also been used to generate a double-knockout rat model in which Cyp3a1 and Cyp3a2 are knocked out (Lu et al, 2017).…”

Section: Drug-metabolizing Enzymesmentioning

confidence: 99%

CRISPR-Cas9: A New Addition to the Drug Metabolism and Disposition Tool Box

et al. 2018

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Clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein 9 (Cas9), i.e., CRISPR-Cas9, has been extensively used as a gene-editing technology during recent years. Unlike earlier technologies for gene editing or gene knockdown, such as zinc finger nucleases and RNA interference, CRISPR-Cas9 is comparably easy to use, affordable, and versatile. Recently, CRISPR-Cas9 has been applied in studies of drug absorption, distribution, metabolism, and excretion (ADME) and for ADME model generation. To date, about 50 papers have been published describing in vitro or in vivo CRISPR-Cas9 gene editing of and-related genes. Twenty of these papers describe gene editing of clinically relevant genes, such as ATP-binding cassette drug transporters and cytochrome P450 drug-metabolizing enzymes. With CRISPR-Cas9, the ADME tool box has been substantially expanded. This new technology allows us to develop better and more predictive in vitro and in vivo ADME models and map previously underexplored genes and gene families. In this mini-review, we give an overview of the CRISPR-Cas9 technology and summarize recent applications of CRISPR-Cas9 within the ADME field. We also speculate about future applications of CRISPR-Cas9 in ADME research.

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“…The CYP2C11-null (knockout) rat model was generated using the CRISPR/Cas9 method to insert a 2 bp GT fragment into exon 6 of CYP2C11 (Wei et al 2018). Rats were cultured at the Laboratory Animal Center of Jiangsu University (SPF grade).…”

Section: Chemicals and Reagentsmentioning

confidence: 99%

Peer Review #1 of "CYP2C11 played a significant role in down-regulating rat blood pressure under the challenge of a high-salt diet (v0.2)"

2019

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Background. Arachidonic acid (AA) is oxidized by cytochrome P450s (CYPs) to form epoxyeicosatrienoic acids (EETs), compounds that modulate ion transport, gene expression, and vasorelaxation. Both CYP2Cs and CYP2Js are involved in kidney EET epoxidation. Methods. In this study, we used a CYP2C11-null rat model to explore the in vivo effects of CYP2C11 on vasorelaxation. For 2 months, CYP2C11-null and wild-type Sprague-Dawley rats were either fed normal lab (0.3% (w/w) sodium chloride) or high-salt (8% (w/w) sodium chloride) diets. Subsequently, an invasive method was used to determine blood pressure. Next, western blots, quantitative PCR, and immunohistochemistry were used to determine renal expression of CYPs involved in AA metabolism.

show abstract

Generation and Characterization of a CYP2C11-Null Rat Model by Using the CRISPR/Cas9 Method

Abstract: CYP2C11 is involved in the metabolism of many drugs in rats. To assess the roles of CYP2C11 in physiology and drug metabolism, a -null rat model was generated using the clustered

Cited by 22 publications

References 45 publications

CYP2C11 played a significant role in down-regulating rat blood pressure under the challenge of a high-salt diet

CYP2C11 played a significant role in down-regulating rat blood pressure under the challenge of a high-salt diet

CRISPR-Cas9: A New Addition to the Drug Metabolism and Disposition Tool Box

Peer Review #1 of "CYP2C11 played a significant role in down-regulating rat blood pressure under the challenge of a high-salt diet (v0.2)"

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