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Genetic deletion of a short fragment of glucokinase in rabbit by CRISPR/Cas9 leading to hyperglycemia and other typical features seen in MODY-2

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Abstract

Glucokinase (GCK) is a key enzyme in glucose sensing and glycemic regulation. In humans, mutations in the GCK gene cause maturity-onset diabetes of the young 2 (MODY-2), a disease that is characterized by an early-onset and persistent hyperglycemia. It is known that Gck knockout (KO) is lethal in mice with Gck KO mice dying within 2 weeks after birth. Therefore, Gck KO mice are not suitable for preclinical study and have limited suitability to study the pathophysiological role of glucokinase in vivo. Here, we report the generation of a novel rabbit with a non-frameshift mutation of GCK gene (GCK-NFS) by cytoplasm microinjection of Cas9 mRNA and gRNA. These GCK-NFS rabbits showed typical features of MODY-2 including hyperglycemia and glucose intolerance with similar survival rate and weight compared to wild-type (WT) rabbits. The diabetic phenotype including pancreatic and renal dysfunction was also found in the F1-generation rabbits, indicating that the genetic modification is germline transmissible. Treatment of GCK-NFS rabbit with glimepiride successfully reduced the fasting blood glucose drastically and improved its islet function. In conclusion, this novel GCK mutant rabbit generated with the CRISPR/Cas9 system mimics most, if not all, histopathological and functional defects seen in MODY-2 patients such as hyperglycemia and will be a valuable rabbit model for preclinical studies and drug screening for diabetes as well as for studying the pathophysiological role of glucokinase.

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Acknowledgements

The authors thank Peiran Hu for excellent technical assistance at the Embryo Engineering Center. This study was financially supported by the National Key Research and Development Program of China Stem Cell and Translational Research (2017YFA0105101). The Strategic Priority Research Program of the Chinese Academy of Sciences (XDA16030501, XDA16030503), Guangdong Province science and technology plan project (2014B020225003). China Postdoctoral Science Foundation Funded Project (2018M641784). China Postdoctoral Science Foundation Funded Project (2018M641784).

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  1. Yuning Song, Tingting Sui and Yuxin Zhang contributed equally to this work.

Authors and Affiliations

  1. Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, 130062, China

    Yuning Song, Tingting Sui, Yuxin Zhang, Yong Wang, Mao Chen, Jichao Deng, Liangxue Lai & Zhanjun Li

  2. Department of Diabetes, Central Clinical School, Pathophysiology of Diabetic Complications Laboratory, Monash University, Melbourne, Australia

    Zhonglin Chai

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  1. Yuning Song
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Song, Y., Sui, T., Zhang, Y. et al. Genetic deletion of a short fragment of glucokinase in rabbit by CRISPR/Cas9 leading to hyperglycemia and other typical features seen in MODY-2. Cell. Mol. Life Sci. 77, 3265–3277 (2020). https://doi.org/10.1007/s00018-019-03354-4

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