Uricase-deficient rat is generated with CRISPR/Cas9 technique

Yun Yu; Nan Zhang; Xianxiang Dong; Nan Fan; Lei Wang; Yuhui Xu; Huan Chen; Weigang Duan

doi:10.7717/peerj.8971

Uricase-deficient rat is generated with CRISPR/Cas9 technique

PeerJ. 2020 Apr 27:8:e8971. doi: 10.7717/peerj.8971. eCollection 2020.

Authors

Yun Yu^#¹, Nan Zhang^#¹, Xianxiang Dong¹, Nan Fan¹, Lei Wang¹, Yuhui Xu¹, Huan Chen², Weigang Duan²

Affiliations

¹ The Department of Pharmacology, School of Basic Medicine, Kunming Medical University, Kunming, Yunnan, China.
² Yunnan Provincial Key Laboratory of Molecular Biology for Sinomedicine, School of Basic Medicine, Yunnan University of Traditional Chinese Medicine, Kunming, Yunnan, China.

^# Contributed equally.

Abstract

Urate oxidase (uricase, Uox) is a big obstacle for scientists to establish stable animal models for studying hyperuricemia and associated disorders. Due to the low survival rate of uricase-deficient mice, we generated a Uox-knockout model animal from Sprague Dawley (SD) rats using the CRISPR/Cas9 technique by deleting exons 2 to 4 of the Uox gene. The uricase-deficient rats were named "Kunming-DY rats", and were apparently healthy with more than a 95% survival up to one year. The male rats' serum uric acid (SUA) increased to 48.3 ± 19.1 µg/ml, significantly higher than those of wild-type rats. Some indexes of the blood fat like total triglyceride, low density lipoprotein, and renal function indexes including blood urea nitrogen and serum creatinine were significantly different from those of wild-type rats, however, all the indexes were close to or in normal ranges. Histological renal changes including mild glomerular/tubular lesions were observed in these uricase-deficient rats. Thus, "Kunming-DY rats" with stable uricase-deficiency were successfully established and are an alternative model animal to study hyperuricemia and associated diseases mimicking human conditions.

Keywords: CRISPR/Cas9 technique; High throughput sequencing; Hyperuricemia; Lipid metabolism; Model animal; Renal function; Uricase-deficiency.

Grants and funding

This work was supported by the Foundation for Scientific Research provided by the Yunnan Province Education Department (2018JS153) and by the National Natural Science Foundation of China (81860162). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.