P and M gene junction is the optimal insertion site in Newcastle disease virus vaccine vector for foreign gene expression

Zhao, Wei; Zhang, Zhenyu; Zsák, László; Yu, Qingzhong

doi:10.1099/vir.0.068437-0

Cited by 56 publications

(50 citation statements)

References 25 publications

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“…Although the immune response to vaccination is influenced by many factors, the expression level of foreign genes is undoubtedly the most important one. To optimize foreign gene expression, previously we and other researchers have investigated the effect of genomic location of the foreign genes on the level of foreign gene expression (Carnero et al, 2009;Zhao & Peeters, 2003;Zhao et al, 2015). The results have shown that the P and M gene junction is the optimal insertion site in NDV vaccine vector for foreign gene expression from an ITU.…”

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confidence: 96%

“…the NP gene. However, the insertion of a foreign gene transcription unit into a promoter-proximal position interfered with NDV replication more seriously than a promoter-distal position, resulting in lower levels of foreign gene expression (Zhao et al, 2015). Besides the insertion site, the size and specific properties of the foreign gene and/or its gene product may also affect NDV replication (Al-Garib et al, 2003;DiNapoli et al, 2007;Gao et al, 2008).…”

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“…Based on the well-accepted 'stop and start' transcription mechanism for non-segmented, negative stranded RNA viruses, NDV is thought to transcribe its genes into mRNAs in a gradient of decreasing mRNA abundance according to the position of the gene relative to the 39 end of the genome (Lamb & Parks, 2013;Samal, 2011). Therefore, the addition of an ITU in the viral genome would attenuate its downstream gene transcription, subsequently may decrease virus replication and the level of foreign gene expression (Krishnamurthy et al, 2000;Skiadopoulos et al, 2000;Wertz et al, 2002;Zhao et al, 2015).…”

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Development of a Newcastle disease virus vector expressing a foreign gene through an internal ribosomal entry site provides direct proof for a sequential transcription mechanism

Zhang

Zhao

et al. 2015

Journal of General Virology

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In the present study, we developed a novel approach for foreign gene expression by Newcastle disease virus (NDV) from a second ORF through an internal ribosomal entry site (IRES). Six NDV LaSota strain-based recombinant viruses vectoring the IRES and a red fluorescence protein (RFP) gene behind the nucleocapsid (NP), phosphoprotein (P), matrix (M), fusion (F), haemagglutinin-neuraminidase (HN) or large polymerase (L) gene ORF were generated using reverse genetics technology. The insertion of the second ORF slightly attenuated virus pathogenicity, but did not affect ability of the virus to grow. Quantitative measurements of RFP expression in virus-infected DF-1 cells revealed that the abundance of viral mRNAs and red fluorescence intensity were positively correlated with the gene order of NDV, 39-NP-P-M-F-HN-L-59, proving the sequential transcription mechanism for NDV. The results herein suggest that the level of foreign gene expression could be regulated by selecting the second ORF insertion site to maximize the efficacy of vaccine and gene therapy. Newcastle disease virus (NDV) is a member of the genus Avulavirus within the subfamily Paramyxovirinae of the family Paramyxoviridae (Lamb et al., 2005). The genome of NDV is a non-segmented, negative-sense ssRNA of 15.2 kb that contains six genes, in the order 39-nucleocapsid (NP)-phosphoprotein (P)-matrix (M)-fusion (F)-haemagglutininneuraminidase (HN)-large polymerase (L)-59, flanked by a 39 UTR leader and 59 trailer Peeters et al., 2000). Two additional proteins (V and W) are produced through editing of the phosphoprotein mRNA (Steward et al., 1993). The genomic RNA is encapsidated with the NP protein and associated with the P and L proteins to form the ribonucleoprotein complex (RNP), which is an active template for viral RNA transcription and replication . Based on their pathogenicity for chickens, NDV strains have been classified into lentogenic (low virulence), mesogenic (intermediate virulence) and velogenic (high virulence) pathotypes (Miller & Koch, 2013). Virulent strains of NDV are the causative agents of Newcastle disease (ND), affecting a wide variety of birds and causing significant economic losses to the poultry industry worldwide (Alexander, 2001). Some lentogenic NDV strains, such as LaSota and B1, have been used as vaccines to protect poultry against ND worldwide.During the late 1990s, reverse genetics technology was developed to rescue infectious NDV from cloned cDNAs Römer-Oberdörfer et al., 1999). Since then, many strains of NDV have been developed as vectors to express foreign genes for vaccine or gene therapy purposes Bukreyev et al., 2005;DiNapoli et al., 2007DiNapoli et al., , 2010 Ge et al., 2007Ge et al., , 2010Hu et al., 2011;Kong et al., 2012; Kortekaas et al., 2010;Niu et al., 2014;Yu et al., 2013;Zhao & Peeters, 2003;Zhao et al., 2014). Usually, the foreign genes are inserted into a non-coding region in the NDV genome as an additional independent transcription unit (ITU) that consists of NDV gene start, the foreign gene and NDV gene...

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Development of a Newcastle disease virus vector expressing a foreign gene through an internal ribosomal entry site provides direct proof for a sequential transcription mechanism

Zhang

Zhao

et al. 2015

Journal of General Virology

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“…For most NDV vaccine vectors, the foreign genes were expressed from an additional transcriptional unit in the NDV genome (Engel-Herbert et al, 2003;Nakaya et al, 2001), which may affect transcription efficiency of the downstream viral genes and virus replication dynamics (Zhao et al, 2015). In this study, we developed a novel approach for the expression of a foreign gene by inserting the reporter gene (GFP) into the M gene transcriptional unit of the thermostable rTS09-C strain to produce a polyprotein GFP-2AUbi-M.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, it is necessary to maximize the foreign gene expression from the NDV vaccine vector by identifying the optimal insertion site. For expression of a foreign gene from an independent transcriptional unit, the gene junction region between the P and M genes has been considered to be the optimal site (Carnero et al, 2009;Zhao & Peeters, 2003;Zhao et al, 2015). However, for expression of a foreign gene from an integrating transcriptional unit, the optimal insertion site has not been studied until now.…”

Section: Discussionmentioning

confidence: 99%

Development of a novel thermostable Newcastle disease virus vaccine vector for expression of a heterologous gene

Wen

Chen

Guo

et al. 2015

Journal of General Virology

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The tumor suppressive effect and apoptotic mechanism of TRAIL gene‐containing recombinant NDV in TRAIL‐resistant colorectal cancer HT‐29 cells and TRAIL‐nonresistant HCT116 cells, with each cell bearing a mouse model

Jung,

An,

Jang

et al. 2023

Cancer Medicine

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BackgroundTRAIL is an anticancer drug that induces cancer cell apoptosis by interacting with death receptors (DRs). However, owing to low cell‐surface expression of DRs, certain colorectal cancer (CRC) cells resist TRAIL‐induced apoptosis. Newcastle disease virus (NDV) infection can elevate DR protein expression in cancer cells, potentially influencing their TRAIL sensitivity. However, the precise mechanism by which NDV infection modulates DR expression and impacts TRAIL sensitivity in cancer cells remains unknown.MethodsHerein, we developed nonpathogenic NDV VG/GA strain‐based recombinant NDV (rNDV) and TRAIL gene‐containing rNDV (rNDV‐TRAIL). We observed that viral infections lead to increased DR and TRAIL expressions and activate signaling proteins involved in intrinsic and extrinsic apoptosis pathways. Experiments were conducted in vitro using TRAIL‐resistant CRC cells (HT‐29) and nonresistant CRC cells (HCT116) and in vivo using relevant mouse models.ResultsrNDV‐TRAIL was found to exhibit better apoptotic efficacy than rNDV in CRC cells. Notably, rNDV‐TRAIL had the stronger cancer cell‐killing effect in TRAIL‐resistant CRC cells. Western blot analyses showed that both rNDV and rNDV‐TRAIL infections activate signaling proteins involved in the intrinsic and extrinsic apoptotic pathways. Notably, rNDV‐TRAIL promotes concurrent intrinsic and extrinsic signal transduction in both HCT‐116 and HT‐29 cells.ConclusionsTherefore, rNDV‐TRAIL infection effectively enhances DR expression in DR‐depressed HT‐29 cells. Moreover, the TRAIL protein expressed by rNDV‐TRAIL effectively interacts with DR, leading to enhanced apoptosis in TRAIL‐resistant HT‐29 cells. Therefore, rNDV‐TRAIL has potential as a promising therapeutic approach for treating TRAIL‐resistant cancers.

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P and M gene junction is the optimal insertion site in Newcastle disease virus vaccine vector for foreign gene expression

Cited by 56 publications

References 25 publications

Development of a Newcastle disease virus vector expressing a foreign gene through an internal ribosomal entry site provides direct proof for a sequential transcription mechanism

Development of a Newcastle disease virus vector expressing a foreign gene through an internal ribosomal entry site provides direct proof for a sequential transcription mechanism

Development of a novel thermostable Newcastle disease virus vaccine vector for expression of a heterologous gene

The tumor suppressive effect and apoptotic mechanism of TRAIL gene‐containing recombinant NDV in TRAIL‐resistant colorectal cancer HT‐29 cells and TRAIL‐nonresistant HCT116 cells, with each cell bearing a mouse model

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