Efficient Mutagenesis of Marek’s Disease Virus-Encoded microRNAs Using a CRISPR/Cas9-Based Gene Editing System

Luo, Jun; Teng, Maikun; Zai, Xusheng; Tang, Na; Zhang, Yaoyao; Mandviwala, Ahmedali; Reddy, V. R.; Baigent, Susan J.; Yao, Yongxiu; Nair, Venugopal

doi:10.3390/v12040466

Cited by 17 publications

(19 citation statements)

References 67 publications

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“…The CRISPR/Cas 9 technique has been widely used to edit the host genes and modify the virus genes [ 31 – 34 ]. This study combined the homology-dependent knock-in and the CRISPR/Cas 9 approach to generate the recombinant virus FA4-EGFP.…”

Section: Discussionmentioning

confidence: 99%

A novel fiber-2-edited live attenuated vaccine candidate against the highly pathogenic serotype 4 fowl adenovirus

Xie

Cao²,

Zhang³

et al. 2021

Vet Res

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Recently, the outbreaks of hydropericardium-hepatitis syndrome (HHS) caused by the highly pathogenic fowl adenovirus serotype 4 (FAdV-4) have resulted in huge economic losses to the poultry industry globally. Although several inactivated or subunit vaccines have been developed against FAdV-4, live-attenuated vaccines for FAdV-4 are rarely reported. In this study, a recombinant virus FA4-EGFP expressing EGFP-Fiber-2 fusion protein was generated by the CRISPR/Cas9 technique. Although FA4-EGFP shows slightly lower replication ability than the wild type (WT) FAdV-4, FA4-EGFP was significantly attenuated in vivo compared with the WT FAdV-4. Chickens infected with FA4-EGFP did not show any clinical signs, and all survived to 14 day post-infection (dpi), whereas those infected with FAdV-4 showed severe clinical signs with HHS and all died at 4 dpi. Besides, the inoculation of FA4-EGFP in chickens provided efficient protection against lethal challenge with FAdV-4. Compared with an inactivated vaccine, FA4-EGFP induced neutralizing antibodies with higher titers earlier. All these data not only provide a live-attenuated vaccine candidate against the highly pathogenic FAdV-4 but also give a potential insertion site for developing FAdV-4-based vaccine vectors for delivering foreign antigens.

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Section: Discussionmentioning

confidence: 99%

A novel fiber-2-edited live attenuated vaccine candidate against the highly pathogenic serotype 4 fowl adenovirus

Xie

Cao²,

Zhang³

et al. 2021

Vet Res

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“…Whether R-LORF8 participates in miR-M2-5p-regulated signaling responsible for cell biological activity requires further study. Along with the development of more powerful mRNA target identification strategies and the applications of CRISPR/Cas9based gene editing technique in research on herpesvirus biology (Bi et al, 2014;Yuen et al, 2015;Bierle et al, 2016;Zhang et al, 2018Zhang et al, , 2019Luo et al, 2020), further breakthrough in the comprehensive understanding of the biological functions of viral miRNAs can be expected to follow.…”

Section: Discussionmentioning

confidence: 99%

Marek’s Disease Virus (Gallid alphaherpesvirus 2)-Encoded miR-M2-5p Simultaneously Promotes Cell Proliferation and Suppresses Apoptosis Through RBM24 and MYOD1-Mediated Signaling Pathways

Zhu

Teng

et al. 2020

Front. Microbiol.

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MicroRNAs (miRNAs) have been demonstrated for their involvement in virus biology and pathogenesis, including functioning as key determinants of virally-induced cancers. As an important oncogenic α-herpesvirus affecting poultry health, Marek's disease virus serotype 1 [Gallid alphaherpesvirus 2 (GaHV-2)] induces rapid-onset T-cell lymphomatous disease commonly referred to as Marek's disease (MD), an excellent biological model for the study of virally-induced cancer in the natural hosts. Previously, we have demonstrated that GaHV-2-encoded miRNAs (especially those within the Meq-cluster) have the potential to act as critical regulators of multiple processes such as virus replication, latency, pathogenesis, and/or oncogenesis. In addition to miR-M4-5p (miR-155 homolog) and miR-M3-5p, we have recently found that miR-M2-5p possibly participate in inducing MD lymphomagenesis. Here, we report the identification of two tumor suppressors, the RNA-binding protein 24 (RBM24) and myogenic differentiation 1 (MYOD1), being two biological targets for miR-M2-5p. Our experiments revealed that as a critical miRNA, miR-M2-5p promotes cell proliferation via regulating the RBM24-mediated p63 overexpression and MYOD1-mediated IGF2 signaling and suppresses apoptosis by targeting the MYOD1-mediated Caspase-3 signaling pathway. Our data present a new strategy of a single viral miRNA exerting dual role to potentially participate in the virallyinduced T-cell lymphomagenesis by simultaneously promoting the cell proliferation and suppressing apoptosis.

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“…Deletion of the pp38 gene from the MDV genomic DNA integrated into the host genomes of virally transformed T-lymphoma cell lines (MDCC-MSB-1 and MDCC-HP8 cells) by the CRISPR/Cas9 system showed an increase in the proliferation of tumor cells, indicating that pp38 is not necessary for the transformation of T-lymphoma cell lines [ 83 ]. Furthermore, a series of virus-encoded microRNAs (miRNAs) in the viral genomes of MDV-1 strain RB-1B or T-lymphoma MSB-1 cell line was successfully mutated by CRISPR/Cas9 system [ 84 , 85 ], providing new important clues for revealing the regulatory roles of viral tiny RNAs in triggering the virally induced T cell lymphomagenesis.…”

Section: Crispr/cas System In Virology Researchmentioning

confidence: 99%

Latest Advances of Virology Research Using CRISPR/Cas9-Based Gene-Editing Technology and Its Application to Vaccine Development

Teng

Yao

Nair

et al. 2021

Viruses

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In recent years, the CRISPR/Cas9-based gene-editing techniques have been well developed and applied widely in several aspects of research in the biological sciences, in many species, including humans, animals, plants, and even in viruses. Modification of the viral genome is crucial for revealing gene function, virus pathogenesis, gene therapy, genetic engineering, and vaccine development. Herein, we have provided a brief review of the different technologies for the modification of the viral genomes. Particularly, we have focused on the recently developed CRISPR/Cas9-based gene-editing system, detailing its origin, functional principles, and touching on its latest achievements in virology research and applications in vaccine development, especially in large DNA viruses of humans and animals. Future prospects of CRISPR/Cas9-based gene-editing technology in virology research, including the potential shortcomings, are also discussed.

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Efficient Mutagenesis of Marek’s Disease Virus-Encoded microRNAs Using a CRISPR/Cas9-Based Gene Editing System

Cited by 17 publications

References 67 publications

A novel fiber-2-edited live attenuated vaccine candidate against the highly pathogenic serotype 4 fowl adenovirus

A novel fiber-2-edited live attenuated vaccine candidate against the highly pathogenic serotype 4 fowl adenovirus

Marek’s Disease Virus (Gallid alphaherpesvirus 2)-Encoded miR-M2-5p Simultaneously Promotes Cell Proliferation and Suppresses Apoptosis Through RBM24 and MYOD1-Mediated Signaling Pathways

Latest Advances of Virology Research Using CRISPR/Cas9-Based Gene-Editing Technology and Its Application to Vaccine Development

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