CRISPR-Cas9-mediated chicken prmt5 gene knockout and its critical role in interferon regulation

CRISPR-Cas9介导的鸡prmt5基因敲除及其在干扰素调控中的关键作用

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作者:Qinghua Zeng, Jingjing Cao, Fei Xie, Lina Zhu, Xiangdong Wu, Xifeng Hu, Zheng Chen, Xiaoqing Chen, Xiangzhi Li, Cheng-Ming Chiang, Huansheng Wu

Abstract

Protein arginine methyltransferase 5 (PRMT5), a type II arginine methyltransferase, controls arginine dimethylation of a variety of substrates. While many papers have reported the function of mammalian PRMT5, it remains unclear how PRMT5 functions in chicken cells. In this study, we found that chicken (ch) PRMT5 is widely expressed in a variety of chicken tissues and is distributed in both the cytoplasm and the nucleus. Ectopic expression of chPRMT5 significantly suppresses chIFN-β activation induced by chMDA5. In addition, a prmt5 gene-deficient DF-1 cell line was constructed using CRISPR/Cas9. In comparison with the wild-type cells, the prmt5-/- DF-1 cells displays normal morphology and maintain proliferative capacity. Luciferase reporter assay and overexpression showed that prmt5-/- DF-1 cells had increased IFN-β production. With identified chicken PRMT5 and CRISPR/Cas9 knockout performed in DF-1 cells, we uncovered a functional link of chPRMT5 in suppression of IFN-β production and interferon-stimulated gene expression.

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