CRISPR-activation screen identified potassium channels for protection against mycotoxins through cell cycle progression and mitochondrial function
Authors:Yulong Tang1,4, Simeng Liao1, Zhuyuan Nie1,2, Guangwei Kuang3, Chunxiao Ji2, Dan Wan1, Liuqin He1, Fengna Li1, Xiangfeng Kong1, Kai Zhan4, Bie Tan2, Xin Wu1 and Yulong Yin1,2
1 Laboratory of Animal Nutritional Physiology and Metabolic Process, Key Laboratory of Agro-ecological Processes in Subtropical Region, National Engineering Laboratory for Pollution Control and Waste Utilization in Livestock and Poultry Production, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China.
2 College of Animal Science and Technology, Hunan Agricultural University, Changsha, China.
3 Hunan Provincial Institute of Animal Drug and Feed Supervision, Changsha, 410006, China.
4 Anhui Province Key Laboratory of Livestock and Poultry Product Safety Engineering, Hefei, 230001, China.
Keywords:
CRISPR-Cas9 screening, zearalenone, potassium channels, cell cycle, mitochondria.
Abbreviations:
NGS – next-generation sequencing,
ROS – reactive oxygen species,
sgRNA – single guide RNA,
ZEA – zearalenone.
Corresponding Author(s):
Conflict of interest statement:
All the authors declare that they have no competing interests.
Please cite this article as:
Yulong Tang, Simeng Liao, Zhuyuan Nie, Guangwei Kuang, Chun-xiao Ji, Dan Wan, Liuqin He, Fengna Li, Xiangfeng Kong, Kai Zhan, Bie Tan, Xin Wu and Yulong Yin (2023). CRISPR-activation screen identified potassium channels for protection against my-cotoxins through cell cycle progression and mitochondrial func-tion. Cell Stress 7(5): 34-45. doi: 10.15698/cst2023.05.279
© 2023 Tang et al. This is an open-access article released under the terms of the Creative Commons Attribution (CC BY) license, which allows the unrestricted use, distribution, and reproduction in any medium, provided the original author and source are acknowledged.
Abstract:
Zearalenone (ZEA) exposure has carcinogenic effects on human and animal health by exhibiting intestinal, hepatic, and renal toxicity. At present, the underlying mechanisms on how ZEA induces apoptosis and damage to tissues still remain unclear. In this study, we aimed to identify genes that modulate the cellular response to ZEA using clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 screening, and further validate novel gene functions to elucidate molecular mechanisms underlying particular biological processes in vivo and in vitro. Two ZEA-resistant cell lines, designated Ov-KCNJ4 and Ov-KCNJ12, were yielded by CRISPR activation screening which had significant changes in ZEA resistance and growth rates. Results showed that ZEA could interact with the cell membrane proteins KCNJ4 and KCNJ12, inducing cell cycle arrest, disruption of DNA replication and base excision repair. Overexpression of KCNJ4 and KCNJ12 was involved in ZEA resistance by regulating cell cycle to neutralize toxicity, sustaining mitochondrial morphology and function via attenuating the damage from oxidative stress in the KCNJ4-mitoKATP pathway. In vivo experiments showed that AAV-KCNJ4 delivery significantly improved ZEA-induced renal impairment and increased antioxidative enzyme activity by improving mitochondrial function. Our findings suggest that increasing potassium channel levels may be a putative therapeutic target for mycotoxin-induced damage.