Generation of a purely clonal defective interfering influenza virus

Abstract

Defective interfering (DI) influenza viruses carry a large deletion in a gene segment that interferes with the replication of infectious virus; thus, such viruses have potential for antiviral therapy. However, because DI viruses cannot replicate autonomously without the aid of an infectious helper virus, clonal DI virus stocks that are not contaminated with helper virus have not yet been generated. To overcome this problem, we used reverse genetics to generate a clonal DI virus with a PB2 DI gene, amplified the clonal DI virus using a cell line stably expressing the PB2 protein, and confirmed its ability to interfere with infectious virus replication in vitro. Thus, our approach is suitable for obtaining purely clonal DI viruses, will contribute to the understanding of DI virus interference mechanisms and can be used to develop DI virus‐based antivirals.

OP7, a novel influenza A virus defective interfering particle: production, purification, and animal experiments demonstrating antiviral potential

“…One approach to avoid the need of UV irradiation may be the production of pure OP7 particles using a cell line that complements the defect in replication of OP7. Such a system was already successfully applied for the generation of conventional DIPs carrying a deletion on Seg1 vRNA using a complementing cell line expressing the corresponding missing protein polymerase basic 2 (Bdeir et al 2019;Yamagata et al 2019). A similar strategy might be used for the production of OP7, for example, by using a cell line expressing the wild-type M1 and/or M2 protein.…”

Defective Interfering Particles with Broad-Acting Antiviral Activity for Dengue, Zika, Yellow Fever, Respiratory Syncytial and SARS-CoV-2 Virus Infection

“…Interestingly, IAV DIPs could also block replication of SARS-CoV-2 in susceptible human cell lines, where inhibition of virus was dependent on janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling ( 12 ). Methods to produce DIPs in the absence of infectious virus have been reported and represent an important achievement as the potential clinical applications of DIPs are advanced ( 17 – 19 ).…”

The Antiviral and Antitumor Effects of Defective Interfering Particles/Genomes and Their Mechanisms

“…Scientists have built an experimental and computational framework for detecting DIPs-associated deletions in the influenza A and B virus, using deep genome sequencing approaches and the precise mechanisms underlying the regulation of DIPs/DI genomes (Alnaji et al, 2019). Reports show that it is now possible to generate pure production of DIPs/DI genomes in the absence of standard virus in vitro (Bdeir et al, 2019; Yamagata et al, 2019). These techniques shed light on the further development of DIPs/DI genome because of the improved safety in their use.…”