Aug 2026· Synthetic and Systems Biotechnology· Vol 17, pp. 41 - 50· 0 citations· 43 references
Medicine
TL;DR
A modular, helper-virus-free and high-efficiency rescue platform based on an orthogonal transcription system utilizing orthogonal promoters and engineered RNA polymerases fused to an mRNA capping enzyme is developed and provides a proof-of-concept methodological framework for the rapid development of vaccine candidates.
Abstract
Recombinant measles virus (rMeV) vectors are promising platforms for vaccine development against emerging infectious diseases due to their safety, stability, and potent immunogenicity. However, conventional rMeV rescue systems frequently exhibit low efficiency, thereby constraining their scalability and throughput. In this study, we developed a modular, helper-virus-free and high-efficiency rescue platform based on an orthogonal transcription system utilizing orthogonal promoters and engineered RNA polymerases fused to an mRNA capping enzyme. This innovative system facilitated robust cytoplasmic manufacture of both genomic and auxiliary components, eliminating the need for helper virus co-infection (such as modified vaccinia virus) and enhancing rescue efficiency by more than 50-fold relative to traditional rescue approaches. Utilizing this technology, we demonstrated the versatility of the platform by successfully generating six rMeV-based vaccine antigen candidates from influenza virus, Pseudomonas aeruginosa, and Brucella spp. All rescued vaccine candidates exhibited stable transgene expression, sustained replication, and strong antigen production. Immunization studies in golden Syrian hamsters verified that the vaccine candidates elicited high titers of neutralizing and antigen-specific antibodies without any observable adverse effects. These results demonstrate that our orthogonal transcription-based platform facilitates the efficient and safe production of rMeV vectors and provides a proof-of-concept methodological framework for the rapid development of vaccine candidates.
By systematically removing host restriction factors, this platform provides a versatile and powerful strategy for accelerating viral propagation, offering a strong foundation for more efficient development and large-scale production of cell culture-based anti-viral vaccines.
Zheng-Mei Xu, Jaemyeong Jeon, Jinsoo Oh et al.· Frontiers in Bioengineering...· 0 citations
It is demonstrated that NTV-ΔF1L-C7L combines high safety with enhanced replication and immunogenicity, supporting its value as a novel vaccine vector and its potential application in controlling the current global monkeypox outbreak.
Jiao Ren, Shi-Yuan Liu, Hang Yuan et al.· Journal of Virology· 0 citations
This review systematically summarizes the evolution, technical framework and optimization strategies of RNA virus reverse genetics, with three prototype viruses covering all major RNA genome types: SARS-CoV-2 (+ssRNA), non-segmented negative-sense Newcastle disease virus (NDV), and segmented negative-sense influenza A virus.
Yu Guo, Ting Xue, Jianhua Wang et al.· Frontiers in Virology· 0 citations
This review comprehensively evaluates the rational design of classical animal herpesvirus vectors, including pseudorabies virus, herpesvirus of turkeys, and feline herpesvirus type 1, providing perspectives on how continuous biotechnological innovations will empower herpesvirus vectors to serve as formidable prophylactic tools against emerging and re-emerging infectious diseases.
Jia-Hui Guo, Chen Mei, Xin-Yao Sun et al.· Frontiers in Microbiology· 0 citations
Classical swine fever virus (CSFV) and pseudorabies virus (PRV) remain significant threats to the swine industry. Although recombinant PRV vectors represent promising platforms for bivalent vaccine development, their application is frequently limited by insufficient expression of heterologous antigens, which may compromise protective efficacy. To address this limitation, we employed a multi-copy expression strategy to enhance CSFV E2 protein levels in a PRV-based vector. Using CRISPR/Cas9-mediated gene editing, we constructed recombinant PRVs expressing one, two, or three copies of the E2 gene by sequential insertion into the gE/gI, tk, and gG loci. In vitro validation demonstrated increased detectable E2 expression in the multi-copy recombinant viruses. Meanwhile, the recombinant viruses maintained virion morphology and replication kinetics comparable to those of the parental strain PRV-GX. Immunogenicity studies in rabbits showed that PRV-3CE2 elicited stronger E2-specific humoral responses and E2-associated cytokine recall responses than PRV-2CE2, while both recombinant viruses induced detectable neutralizing activity. In challenge experiments, although vaccination did not completely prevent febrile responses following CSFV challenge, both PRV-2CE2 and PRV-3CE2 reduced CSFV RNA loads in blood compared with the mock group, suggesting partial protective efficacy. Additionally, both PRV-2CE2 and PRV-3CE2 provided complete protection against lethal PRV challenge in rabbits. These findings suggest that multi-copy E2 expression enhances E2-associated immunogenicity and supports further optimization and evaluation of PRV-3CE2 as a PRV-CSFV bivalent vaccine candidate.
Xianfei Shang, Hui Zhao, Yufeng He et al.· Veterinary Microbiology· 0 citations
Multi pathogen vaccines have emerged as a promising strategy to improve vaccine coverage, simplify immunization logistics, and address overlapping global health threats. However, few vaccine platforms have demonstrated robust and durable immune responses against pathogens spanning distinct biological classes. Here, we developed and evaluated a viral-vectored multipathogen vaccine platform based on a heterologous prime–boost regimen combining the highly attenuated vaccinia virus LC16m8Δ (m8Δ) and adeno-associated virus serotype 1 (AAV1). As a preclinical proof-of-concept, antigen-specific immune responses against Plasmodium falciparum and SARS-CoV-2 (Omicron variant) were evaluated, while neutralizing activity against mpox virus was assessed to examine the cross-reactive immunity conferred by the vaccinia virus vector. Immunogenicity, protective efficacy, and transmission-blocking activity were evaluated in murine models and mosquito feeding assays. The heterologous m8Δ prime/AAV1 boost regimen induced robust and long-lasting antigen-specific antibody responses that were maintained for up to 32 weeks. Complete sterile protection against transgenic Plasmodium berghei sporozoite challenge was achieved, together with greater than 90% transmission-blocking efficacy in mosquito feeding assays. In parallel, potent neutralizing antibody responses against SARS-CoV-2 Omicron and cross-neutralizing activity against mpox virus were observed. These findings demonstrate the feasibility of the m8Δ/AAV1 platform as a versatile viral-vectored multipathogen vaccine platform capable of integrating protozoan and viral antigens, providing a rationale for further preclinical optimization and clinical evaluation of next generation multipathogen vaccines targeting both endemic and emerging infectious diseases.
Yuna Sato, Yutaro Yamamoto, A. Hasyim et al.· Frontiers in Immunology· 0 citations
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