CRISPR/Cas9 genome editing technology significantly accelerated herpes simplex virus research

Area of Science:

• Virology and Molecular Biology
• Gene Therapy
• Genome Editing Technologies

Background:

• Herpes simplex viruses (HSVs) are significant pathogens with large genomes, making them suitable for gene therapy applications.
• Traditional methods for constructing recombinant HSVs, relying on DNA homology-dependent repair (HDR) and plaque assays, are inefficient and time-consuming.

Purpose of the Study:

• To review the latest advancements in CRISPR/Cas9 genome editing technology.
• To explore the molecular mechanisms of CRISPR/Cas9.
• To summarize the recent applications of CRISPR/Cas9 in engineering herpes simplex viruses (HSVs).

Main Methods:

• Application of clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9 (CRISPR/Cas9) for precise and efficient genome editing.
• Integration of CRISPR/Cas9 with flow cytometric technology for automated HSV recombination.
• Review of existing literature on CRISPR/Cas9 mediated HSV genome modification.

Main Results:

• CRISPR/Cas9 technology has exponentially increased the efficiency of HSV homology-dependent repair (HDR) by 10,000 to 1,000,000 times.
• The combination of CRISPR/Cas9 and flow cytometry has made HSV recombination nearly automatic.
• CRISPR/Cas9 enables rapid and precise genome editing in HSVs.

Conclusions:

• CRISPR/Cas9 technology represents a significant breakthrough in constructing recombinant HSVs.
• This advancement is poised to have a substantial impact on virus research and the development of gene therapies.
• The review highlights the transformative potential of CRISPR/Cas9 in virology and genetic engineering.