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Progress and Applications of Reverse Genetics in Virology

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Special Issue Editor

Dr. Yang Liu

E-Mail Website
Guest Editor

Institute of Infectious Diseases, Shenzhen Bay Laboratory, Shenzhen, China
Interests: investigating the genetic evolution of important human pathogenic viruses; the pathogenesis of severe viral diseases and the molecular mechanisms of viral infection and transmission; developing novel viral tools and antiviral strategies

Special Issue Information

Dear Colleagues,

Reverse genetics, diverging from traditional forward genetics, has reshaped our understanding of genotype–phenotype relationships. By directly altering genetic sequences to achieve a desired phenotype, it surpasses the standard approach based on genetic variance. Modern molecular techniques and genome sequencing have expanded the reach and applicability of reverse genetics.

Viruses, due to their simplified genomic structures, serve as prime subjects for reverse genetics. Their readily modifiable genomes have led to significant advancements in viral biology, unearthing gene functions, optimizing microbiological resources, reviving rare viral strains, and aiding in vaccine development.

For this Special Issue, "Progress and Applications of Reverse Genetics in Virology", we invite the submission of original research and review articles focused on new technologies and novel approaches for the implications of reverse genetics, including (but not limited to) dsDNA virus, ssDNA virus, dsRNA virus, +ssRNA virus, -ssRNA virus, Ambisense RNA virus, etc.

Dr. Yang Liu
Guest Editor

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Published Papers (3 papers)

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Research

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15 pages, 3056 KiB

Open AccessArticle

Amino Acids at Positions 156 and 332 in the E Protein of the West Nile Virus Subtype Kunjin Virus Classical Strain OR393 Are Involved in Plaque Size, Growth, and Pathogenicity in Mice

by Shigeru Tajima, Hideki Ebihara and Chang-Kweng Lim

Viruses 2024, 16(8), 1237; https://doi.org/10.3390/v16081237 - 1 Aug 2024

Cited by 1 | Viewed by 814 | Correction

Abstract

The West Nile virus (WNV) subtype Kunjin virus (WNV_KUN) is endemic to Australia. Here, we characterized the classical WNV_KUN strain, OR393. The original OR393 strain contained two types of viruses: small plaque-forming virus (SP) and large plaque-forming virus (LP). The amino acid residues at positions 156 and 332 in the E protein (E¹⁵⁶ and E³³²) of SP were Ser and Lys (E^156S/332K), respectively, whereas those in LP were Phe and Thr (E^156F/332T). SP grew slightly faster than LP in vitro. The E protein of SP was N-glycosylated, whereas that of LP was not. Analysis using two recombinant single-mutant LP viruses, rKUNV-LP-E^F156S and rKUNV-LP-E^T332K, indicated that E^156S enlarged plaques formed by LP, but E^332K potently reduced them, regardless of the amino acid at E¹⁵⁶. rKUNV-LP-E^F156S showed significantly higher neuroinvasive ability than LP, SP, and rKUNV-LP-E^T332K. Our results indicate that the low-pathogenic classical WNV_KUN can easily change its pathogenicity through only a few amino acid substitutions in the E protein. It was also found that Phe at E¹⁵⁶ of the rKUNV-LP-E^T332K was easily changed to Ser during replication in vitro and in vivo, suggesting that E^156S is advantageous for the propagation of WNV_KUN in mammalian cells. Full article

(This article belongs to the Special Issue Progress and Applications of Reverse Genetics in Virology)

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Review

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16 pages, 774 KiB

Open AccessReview

The Role of Noncoding RNA in the Transmission and Pathogenicity of Flaviviruses

by Xianwen Zhang, Yuhan Li, Yingyi Cao, Ying Wu and Gong Cheng

Viruses 2024, 16(2), 242; https://doi.org/10.3390/v16020242 - 2 Feb 2024

Cited by 1 | Viewed by 2534

Abstract

Noncoding RNAs (ncRNAs) constitute a class of RNA molecules that lack protein-coding capacity. ncRNAs frequently modulate gene expression through specific interactions with target proteins or messenger RNAs, thereby playing integral roles in a wide array of cellular processes. The Flavivirus genus comprises several significant members, such as dengue virus (DENV), Zika virus (ZIKV), and yellow fever virus (YFV), which have caused global outbreaks, resulting in high morbidity and mortality in human populations. The life cycle of arthropod-borne flaviviruses encompasses their transmission between hematophagous insect vectors and mammalian hosts. During this process, a complex three-way interplay occurs among the pathogen, vector, and host, with ncRNAs exerting a critical regulatory influence. ncRNAs not only constitute a crucial regulatory mechanism that has emerged from the coevolution of viruses and their hosts but also hold potential as antiviral targets for controlling flavivirus epidemics. This review introduces the biogenesis of flavivirus-derived ncRNAs and summarizes the regulatory roles of ncRNAs in viral replication, vector-mediated viral transmission, antiviral innate immunity, and viral pathogenicity. A profound comprehension of the interplay between ncRNAs and flaviviruses will help formulate efficacious prophylactic and therapeutic strategies against flavivirus-related diseases. Full article

(This article belongs to the Special Issue Progress and Applications of Reverse Genetics in Virology)

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