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CRISPR-Cas9 Mediated RNase L Knockout Regulates Cellular Function of PK-15 Cells and Increases PRV Replication

文献类型： 外文期刊

作者： Sui, Chao 1 ; Jiang, Dandan 2 ; Wu, Xiangju 2 ; Cong, Xiaoyan 2 ; Li, Feng 3 ; Shang, Yingli 1 ; Wang, Jinqiu 4 ; Liu, Si 1 ;

作者机构： 1.Shandong Agr Univ, Coll Anim Sci & Technol, Shandong Prov Key Lab Anim Biotechnol & Dis Contr, Tai An 271018, Shandong, Peoples R China

2.Shandong Acad Agr Sci, Inst Anim Sci & Vet Med, Shandong Key Lab Anim Dis Control & Breeding, Sangyuan Rd 8, Jinan 250100, Shandong, Peoples R China

3.South Dakota State Univ, Dept Vet & Biomed Sci, Dept Biol & Microbiol, Brookings, SD 57007 USA

4.Beijing Vocat Coll Agr, Dept Anim Husb & Vet Med, 5 Daotiannanli Rd, Beijing 102442, Peoples R China

5.Qingdao Agr Univ, Coll Vet Med, Qingdao 266109, Peoples R China

6.Shandong Normal Univ, Coll Life Sci, Jinan 250014, Shandong, Peoples R China

期刊名称：BIOMED RESEARCH INTERNATIONAL （ 影响因子：3.411； 五年影响因子：3.62 ）

ISSN： 2314-6133

年卷期： 2019 年

页码：

收录情况： SCI

摘要： Ribonuclease L (RNase L) is an important antiviral endoribonuclease regulated by type I IFN. RNase L is activated by viral infection and dsRNA. Because the role of swine RNase L (sRNase L) is not fully understood, in this study, we generated a sRNase L knockout PK-15 (KO-PK) cell line through the CRISPR/Cas9 gene editing system to evaluate the function of sRNase L. After transfection with CRISPR-Cas9 followed by selection using puromycin, sRNase L knockout in PK-15 cells was further validated by agarose gel electrophoresis, DNA sequencing, and Western blotting. The sRNase L KO-PK cells failed to trigger RNA degradation and induced less apoptosis than the parental PK-15 cells after transfected with poly (I: C). Furthermore, the levels of ISGs mRNA in sRNase L KO-PK cells were higher than those in the parental PK-15 cells after treated with poly (I: C). Finally, both wild type and attenuated pseudorabies viruses (PRV) replicated more efficiently in sRNase L KO-PK cells than the parental PK-15 cells. Taken together, these findings suggest that sRNase L has multiple biological functions including cellular single-stranded RNA degradation, induction of apoptosis, downregulation of transcript levels of ISGs, and antiviral activity against PRV. The sRNase L KO-PK cell line will be a valuable tool for studying functions of sRNase L as well as for producing PRV attenuated vaccine.

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