2. Academic Validation
  3. Genetic determinants of efficacy of antiviral drugs revealed by genome-wide CRISPR screens

Genetic determinants of efficacy of antiviral drugs revealed by genome-wide CRISPR screens

  • Antiviral Res. 2025 Dec:244:106309. doi: 10.1016/j.antiviral.2025.106309.
Wei Jiang 1,  Ailing Yang 2,  Jingchuan Ma 2,  Dawei Lv 3,  Mingxian Liu 2,  Chao Wang 2,  Shuo Chen 1,  Huiling Fang 2,  Yankai Chu 2,  Zhengjin He 2,  Wenrui Li 1,  Yucheng Liu 1,  Yun Zhao 4,  Zhaocai Zhou 5,  Gang Long 6,  Hai Jiang 7
Affiliations

Affiliations

  • 1 Key Laboratory of Systems Health Science of Zhejiang Province, School of Life Science, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, 310024, China.
  • 2 Key Laboratory of RNA Innovation, Science and Engineering, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, 200031, China.
  • 3 Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, Shanghai Medical College, Biosafety Level 3 Laboratory, Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai, China.
  • 4 Key Laboratory of Systems Health Science of Zhejiang Province, School of Life Science, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, 310024, China; Key Laboratory of RNA Innovation, Science and Engineering, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, 200031, China; School of Life Science and Technology, ShanghaiTech University, Shanghai, 201210, China.
  • 5 State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, 200438, China; School of Life Science and Technology, ShanghaiTech University, Shanghai, 201210, China.
  • 6 Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, Shanghai Medical College, Biosafety Level 3 Laboratory, Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai, China. Electronic address: [email protected].
  • 7 Key Laboratory of Systems Health Science of Zhejiang Province, School of Life Science, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, 310024, China; Key Laboratory of RNA Innovation, Science and Engineering, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, 200031, China. Electronic address: [email protected].
PMID: 41260510 DOI: 10.1016/j.antiviral.2025.106309
Abstract

Nucleoside and nucleobase analog Antiviral drugs are pivotal in Antiviral therapy, but comprehensive methods to understand their cellular response mechanisms and genetic regulators are still lacking. Here, we show that Eμ-Myc; Arf-/- mouse lymphoma cells, which are highly apoptosis-prone, enabled genome-wide CRISPR-Cas9 screening on such drugs to identify genes that modulate their efficacy. Using retroviral sgRNA libraries and MAGeCK analysis, we uncovered key regulators of drug transport, activation, and inactivation for these drugs. For ribavirin, Adenosine Kinase (ADK) and adenylsuccinate synthase (ADSS) were critical for nucleotide metabolism and bioactivation. Remdesivir uptake and activation depended on the transporter SLC29A3 and phosphoamidase HINT1, whereas favipiravir resistance was linked to NT5C2-mediated dephosphorylation. Viral replication assays in Huh7 cells validated that knockout of SLC29A3, HINT1, or NT5C2 significantly altered Antiviral efficacy. This study delineates the genetic network governing nucleotide analog response, providing mechanistic insights and potential biomarkers for personalized Antiviral therapy.

Keywords

Antiviral efficacy; Biomarkers; CRISPR screening; Drug response mechanism; Genetic regulators; Nucleoside and nucleobase analog antiviral drugs.

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