Jun 2026· Frontiers in Genome Editing· Vol 8· 0 citations· 59 references
Medicine
TL;DR
Recent advances in CRISPR-based strategies for influenza prevention and control are summarized and it is concluded that with continued technological refinement and careful translational development, CRISPR may become a versatile tool for influenza prevention, diagnosis, and therapy.
Abstract
Influenza A virus (IAV) and influenza B virus (IBV) remain major global public health threats because of their rapid antigenic evolution and efficient human-to-human transmission. In contrast, influenza C virus (ICV) and influenza D virus (IDV) generally exhibit narrower host ranges and milder pathogenicity, yet their potential for interspecies transmission and zoonotic spillover still warrants attention. Conventional prevention strategies, such as inactivated and live-attenuated vaccines, suffer from prolonged development timelines and diminished efficacy against rapidly evolving viral strains. However, antiviral drugs are increasingly limited by the rapid emergence of drug-resistant variants. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) gene-editing technology has emerged as a promising platform for influenza prevention and control owing to its programmability and precise targeting capability. In this paper, we summarize recent advances in CRISPR-based strategies for influenza prevention and control. The RNA-targeting CRISPR-associated protein 13 (Cas13) system can recognize conserved viral RNA sequences and suppress replication across influenza subtypes, whereas the DNA-targeting CRISPR-associated protein 9 (Cas9) system can edit host susceptibility genes and thereby reduce cellular permissiveness to infection. In addition, lipid nanoparticle (LNP)-based delivery systems have become important tools for improving the in vivo delivery and expression of CRISPR components by enhancing targeting efficiency and reducing immunogenicity. CRISPR-based diagnostics, such as Specific High-sensitivity Enzymatic Reporter unLOCKing (SHERLOCK), further expand the clinical utility of this technology by enabling rapid and sensitive detection of influenza viruses. Despite these advances, substantial challenges remain, including delivery inefficiency, off-target activity, long-term safety concerns, and the risk of viral escape. With continued technological refinement and careful translational development, CRISPR may become a versatile tool for influenza prevention, diagnosis, and therapy.
Highly pathogenic avian influenza viruses (HPAIVs) continue to cause substantial disease in birds and mammals, with repeated H5N1 spillovers highlighting the need for broadly protective antiviral strategies. Here, we develop a programmable RNA-targeting antiviral platform based on RfxCas13d and evaluate its activity in avian cells. Screening of five Cas13 orthologs in chicken DF1 fibroblasts revealed RfxCas13d as the most potent and well tolerated effector. Virus-specific CRISPR RNAs (crRNAs) targeting conserved regions of positive- and negative-sense influenza RNA were tested against A/WSN/033 (H1N1) and multiple HPAIV isolates, including a member of clade 2.3.4.4b H5N1. Targeting positive-sense RNA conferred superior influenza inhibitory activity and was further enhanced by multiplexed crRNA expression. These findings establish RfxCas13d as a versatile RNA-guided antiviral platform and provide a foundation for cross-strain influenza control through conserved RNA targeting.
Sudip Dhakal, Aaron J. Smith, Emiliana Weiss et al.· npj Antimicrobials and Resis...· 0 citations
Recombinant herpesvirus vectors have emerged as highly potent platforms for next-generation vaccine development, distinguished by their genomic capacity and their unique ability to elicit durable, long-lasting immunity through persistent latent infections. In this review, we trace the evolutionary trajectory of herpesvirus vector engineering—transitioning from traditional homologous recombination to advanced bacterial artificial chromosome (BAC) systems and precise, scarless CRISPR/Cas9 gene editing. We comprehensively evaluate the rational design of classical animal herpesvirus vectors, including pseudorabies virus (PRV), herpesvirus of turkeys (HVT), and feline herpesvirus type 1 (FHV-1). Specifically, we highlight their distinct advantages in molecular attenuation, the optimization of non-essential insertion loci, and the application of tissue-specific promoters for multiplexed antigen presentation. Furthermore, we discuss the strategic deployment of multivalent herpesvirus vaccines within the “One Health” framework, emphasizing their critical role in simplifying immunization protocols and interrupting the transmission chains of zoonotic diseases. Finally, we address the prevailing translational bottlenecks, including scalable manufacturing challenges and stringent regulatory frameworks regarding environmental release, providing perspectives on how continuous biotechnological innovations will empower herpesvirus vectors to serve as formidable prophylactic tools against emerging and re-emerging infectious diseases.
Jiahui Guo, Chen Mei, Xinyao Sun et al.· Frontiers in Microbiology· 0 citations
Viral infectious diseases in livestock and poultry cause substantial economic losses worldwide and pose persistent zoonotic threats to public health. Elucidating virus-host interactions is essential for understanding viral pathogenesis and developing effective control strategies. In recent years, genome-wide CRISPR/Cas9 functional screening has emerged as a powerful and unbiased high-throughput approach for identifying host dependency and restriction factors. Enabled by the development of species-specific sgRNA libraries, this technology has significantly advanced research in veterinary virology. In this review, we systematically summarize recent progress in CRISPR/Cas9-based screening studies of major livestock and poultry viruses, including foot-and-mouth disease virus (FMDV), swine enteric coronaviruses, African swine fever virus (ASFV), porcine reproductive and respiratory syndrome virus (PRRSV), avian leukosis virus (ALV), and other zoonotic pathogens. We highlight key host factors involved in viral entry, replication, and egress, and integrate these findings to delineate conserved cross-viral dependency networks, such as sialic acid biosynthesis, endosomal–lysosomal trafficking, double-membrane vesicle (DMV) formation, and interferon signaling pathways. Furthermore, we discuss the translational potential of these genomic discoveries for practical agricultural applications, particularly in the development of host-targeted broad-spectrum antivirals and the generation of disease-resistant livestock (e.g., receptor-edited pigs and chickens) through precise genome editing. Finally, we outline future perspectives, including the integration of single-cell transcriptomics and in vivo validation, thereby providing a comprehensive framework for advancing disease control and sustainable breeding in animal agriculture.
This review critically evaluates CRISPR's role in RNA editing, emphasizing its importance for functional genomics and development of recombinant vaccines, andTranslational challenges are critically discussed, including off-target effects, delivery limitations, and ethical issues.
Mahmood Ul Hasan, M. Azhar, Noor-Ul-Saba et al.· Journal of Virological Metho...· 0 citations
Influenza virus remains a major global public health threat due to its high transmissibility, ability to cause severe disease, and pandemic potential. Although vaccination is the main strategy for reducing the burden of influenza, it has notable limitations. Therefore, novel therapeutic and immunomodulatory approaches are urgently needed. Here, using plasmid-driven reverse genetics, we engineered a defective recombinant influenza virus encoding interleukin 7 (Flu:IL-7), to investigate the role of IL-7 in influenza pathogenesis and immunomodulation in a murine model. Our findings demonstrate that Flu:IL-7 is safe and effectively attenuates disease severity caused by a wild-type replicative influenza virus (PR8), improving clinical recovery and reducing disease-associated morbidity. Notably, local expression of IL-7 through the Flu:IL-7 vector at the time of PR8 infection significantly reduced the severity of secondary pneumococcal pneumonia. These protective effects were associated with earlier and more coordinated pulmonary immune response, characterized by enhanced activation and expansion of innate and adaptive immune cells, early formation of iBALT-like aggregates, and reduced tissue damage. Collectively, our findings suggest that transient local IL-7 expression temporally reprograms the pulmonary immune response, promoting early antiviral immunity followed by timely resolution of inflammation. In summary, Flu:IL-7 provides a useful experimental model for investigating the immunomodulatory role of IL-7 during influenza infection. Our findings support transient local IL-7 expression as a promising host-directed strategy to improve disease outcome and reduce influenza-associated complications. These concepts may also be applicable to other diseases in which modulation of the local immune response is desirable, including lung cancer and COVID-19.
Sarah Messias, Ianca Évelin Silva de Paula, K. Cardoso et al.· Immunology Letters· 0 citations
Highly pathogenic RNA viruses, such as Ebola, SARS-CoV-2, and influenza, cause severe disease in humans. High mutation rates, which enable RNA viruses to evade immunity and escape antivirals, and their ability to spread from animals to humans and cause pandemics and outbreaks, make RNA viruses significant threats to public health. Diseases caused by Ebola, SARS-CoV-2, and influenza are prevented and treated with only a limited number of approved antiviral drugs, the effectiveness of which is limited by mutations in the viral targets. It is crucial to understand the structural determinants, molecular mechanisms, and host interactions of pathogenic RNA viruses to develop effective antiviral strategies. In this review, we discuss selected RNA viruses, focusing on the structure of their RNA polymerases and interactions with host factors during the different stages of the viral lifecycle, as well as the traditional antivirals targeting these structures and pathways. Furthermore, emerging concepts such as liquid–liquid phase separation and biomolecular condensates, and novel promising antiviral strategies are discussed. Understanding shared and distinct structures, molecular mechanisms, and host interactions across highly pathogenic RNA viruses enables the discovery of new and more effective antiviral strategies, ultimately improving clinical outcomes against evolving RNA viruses.
Hana Krnjić, Adna Hrapović, Aiša Galijatović et al.· Viruses· 0 citations