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CRISPR-Cas9 system-driven site-specific selection pressure on Herpes simplex virus genomes
Zhihua Li1, Yanwei Bi1, Hongjian Xiao1
1Institute of Medical Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, 935 Jiaoling Road, Wuhua District, Kunming, Yunnan, 650118 China; Yunnan Key Laboratory of Vaccine Research and Development of Severe Infectious Disease, Kunming, Yunnan, China; Centre for Tuberculosis Research, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Virus Research
|March 11, 2017
Summary
The CRISPR-Cas9 system effectively inhibits herpes simplex virus 1 (HSV-1) replication by targeting viral genomes. This method enables precise virus selection and purification without antibiotics or reporter genes.
Area of Science:
- Molecular Biology
- Virology
- Gene Editing Technologies
Background:
- The CRISPR-Cas9 system is a versatile tool for genome editing in eukaryotic cells.
- Viral replication poses significant challenges in research and therapeutic applications.
Purpose of the Study:
- To investigate the efficacy of the CRISPR-Cas9 system for inhibiting herpes simplex virus 1 (HSV-1) replication.
- To explore the potential of CRISPR-Cas9 for specific viral genome selection and purification.
Main Methods:
- Utilized a guide RNA (gRNA) designed to target specific sequences within the HSV-1 genome.
- Employed both wild-type Cas9 protein and Cas9 nickase (D10A) for viral genome targeting.
- Applied CRISPR-Cas9 to a mixed viral pool to assess selection capabilities.
Main Results:
- Demonstrated significant inhibition of HSV-1 replication using the CRISPR-Cas9 system.
- Showcased the system's ability to distinguish between different viral genome sequences with single nucleotide specificity.
- Observed a significant shift in viral proportions within a mixed pool, indicating effective selection pressure.
Conclusions:
- The CRISPR-Cas9 system offers a novel approach to inhibit viral replication and select specific viral genomes.
- This method provides a potentially faster alternative to traditional techniques for screening and purifying mutant viruses, avoiding the need for antibiotics or reporter genes.
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