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Herpes Simplex Virus (HSV) is an enveloped, large, double-stranded DNA virus belonging to the Herpesviridae family. The HSV genome ranges from 150 to 240 kb, encoding approximately 90 viral proteins. The virion diameter is roughly 180 nm, with a 125 nm icosahedral capsid. HSV can infect multiple mammalian cell types, particularly neurons and epithelial cells, with high efficiency. Based on replication capability and application, HSV vectors can be engineered as replication-competent oncolytic viruses (oHSVs) or replication-defective gene therapy vectors, offering broad applications in tumor therapy and gene delivery.
Figure 1. The HSV virion. (Zhu S, et al)
Oncolytic HSVs are genetically engineered viruses that selectively replicate in tumor cells, thereby infecting and killing malignant tissues. Advantages of HSV as a therapeutic vector include a broad host range, high infection efficiency, short replication cycle, large genome capacity, and relatively low pathogenicity. Additionally, antiviral drugs such as acyclovir and ganciclovir can effectively inhibit HSV replication, ensuring safety during laboratory manipulation and downstream applications. Consequently, HSV-1 has become a widely studied platform for oncolytic virus research and clinical development.
The HSV genome is complex, consisting of two unique sequence regions, the long (UL) and short (US) regions, flanked by inverted repeat sequences (TRL-IRL and IRs-TRs). The "a" sequence is located at the junction between IRL and IRs, critical for viral recombination and replication. Approximately half of the HSV genome contains non-essential genes, allowing insertion, deletion, or regulation of exogenous sequences, making HSV an ideal vector system.
Leveraging advanced viral engineering capabilities and extensive HSV genetics expertise, Creative Biogene provides comprehensive HSV modification and production services. Our platform covers recombinant virus construction, viral rescue, amplification, and high-purity virus preparation, catering to research and preclinical needs. Through close collaboration with clients, we design optimal virus construction strategies to achieve flexible and efficient HSV modification.
In oHSV construction, we can perform targeted gene knockouts or insertions in HSV-1. For instance, ICP34.5 (RL1), ICP47, US11, and TK can be selectively deleted to enhance tumor selectivity and safety. Exogenous genes such as GM-CSF or IL-12 can be inserted at specific loci to augment immune responses and anti-tumor activity. Creative Biogene supports a range of HSV-1 strains, including KOS, F, and H129, allowing selection of the most suitable viral background for each project.
Several oHSV engineering strategies have been successfully implemented on our platform. Examples include T-Vec, which has a double ICP34.5 deletion and GM-CSF insertion along with ICP47 deletion; G47Δ, with ICP34.5 deletion, ICP47 deletion, ICP6 reading frame disruption, and LACZ insertion; and M032, featuring double ICP34.5 deletion with IL-12 insertion. Additional strategies include tissue-specific miRNA regulation, UL37 mutations, and multi-gene insertions, showcasing the flexibility and customizability of HSV engineering.
Creative Biogene's HSV service platform offers end-to-end solutions, from client consultation to recombinant virus construction, purification, and quality control. Typical workflow includes:
1Requirement Analysis and Planning: Based on client-provided sequences, target viral strains, and functional objectives, we design personalized HSV modification plans and perform feasibility assessment.
2Targeted Vector Construction: High-fidelity PCR and enzymatic verification are used to construct recombinant vectors, ensuring sequence integrity for insertions, deletions, or regulatory elements.
3Virus Recombination, Plaque Picking, and Purification: Recombinant vectors are co-transfected with HSV genome into packaging cells, followed by virus recombination and plaque selection. Multiple rounds of purification yield high-titer, low-diversity recombinant viruses.
4Molecular Verification and Amplification: Viral genome integrity and modification accuracy are confirmed by PCR, sequencing, and restriction analysis. Viruses are then amplified and purified to meet downstream experimental needs.
Throughout the process, our team uses high-precision quantification to determine viral titers (PFU), genome integrity, and insertion efficiency. Clients receive a comprehensive report including sequencing traces, gel images, restriction maps, and COA files, ensuring transparent and traceable quality control.
Creative Biogene HSV services support diverse applications:
Our advantages include rapid response to client needs, professional experimental design, comprehensive technical support, high-precision virus engineering, and rigorous quality control. Real-time project tracking and detailed reporting ensure transparency and reliability.
Creative Biogene's HSV service platform combines deep virology knowledge, advanced genetic engineering technologies, and a robust quality control system. Whether for oncolytic virus development, gene delivery, neural tracing, or multi-gene engineering, we provide high-precision, efficient, and customizable HSV solutions, accelerating research progress and enabling translational applications. Contact our expert team to design tailored HSV construction and recombinant virus solutions, ensuring efficient, safe, and controlled HSV experimentation.
Q1: Is HSV modification safe? Does exogenous gene insertion affect viral replication?
A: Creative Biogene employs optimized HSV-1 and HSV-2 strains with controlled gene deletions and insertions, ensuring safe laboratory handling and downstream applications. By targeting non-essential regions and regulatory sequences, we preserve viral replication while ensuring functional expression of inserted genes.
Q2: How is viral titer ensured? How to request custom HSV services?
A: Standardized packaging cell systems and high-precision PFU measurements guarantee high-titer virus production, accompanied by verification reports for client reference. Clients provide target sequences or gene information, and we offer full support for sequence optimization, vector design, and virus construction, fulfilling research and preclinical development needs.
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