Increased virus replication in mammalian cells by blocking intracellular innate defense responses

Vries, Walter Timo de; Haasnoot, Joost; Velden, J. van der; Montfort, Thijs van; Zorgdrager, Fokla; Paxton, William A.; Cornelissen, Marion; Kuppeveld, Frank J. M. van; Haan, Peter; Berkhout, Ben

doi:10.1038/gt.2008.12

Cited by 48 publications

(31 citation statements)

References 51 publications

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“…We therefore expected that inhibition of the RNAi pathway during vector production would lead to a (partial) repair of the titers. To test this, we used different approaches to abort the RNAi mechanism by co-transfection of the following: an excess luciferase reporter with the corresponding shRNA target sequence (Luc) as RNAi target decoy; an excess shRNAs (a 5xshRNA plasmid encoding five shRNAs) to saturate the RNAi machinery; a plasmid encoding VA RNA of Adenovirus as Dicer inhibitor (Andersson et al 2005); and a plasmid encoding the RNAi suppressor protein VP35 of Ebola virus (Haasnoot et al 2007a;de Vries et al 2008), siRNAs against Dicer (Paddison et al 2002), or an shRNA against Drosha. Furthermore, we tested whether overexpression of the chromosome region maintenance 1 (CRM1) protein (Popa et al 2002), which is involved in the nuclear export of unspliced and partially spliced viral RNAs, could improve production of viral particles and thereby the titer (Wodrich and Krausslich 2001;Rawlinson et al 2009).…”

Section: Inhibition Of the Rnai Pathway To Increase The Lentiviral Vementioning

confidence: 99%

“…This reporter was also used to produce an RNAi decoy for CMV-miRNA S because it also encodes a target for this miRNA. The plasmid expressing the Adenovirus VA RNAs (pVA RNAs) (Andersson et al 2005), the Ebola VP35 protein (Haasnoot et al 2007a;de Vries et al 2008), the CRM1 co-factor (Popa et al 2002), the siRNA against Dicer, and the shRNA against Luciferase (Paddison et al 2002) have been described elsewhere.…”

Section: Dna Constructsmentioning

confidence: 99%

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Titers of lentiviral vectors encoding shRNAs and miRNAs are reduced by different mechanisms that require distinct repair strategies

Liu¹,

Vink²,

Westerink³

et al. 2010

RNA

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RNAi-based gene therapy is a powerful approach to treat viral infections because of its high efficiency and sequence specificity. The HIV-1-based lentiviral vector system is suitable for the delivery of RNAi inducers to HIV-1 susceptible cells due to its ability to transduce nondividing cells, including hematopoietic stem cells, and its ability for stable transgene delivery into the host cell genome. However, the presence of anti-HIV short hairpin RNA (shRNA) and microRNA (miRNA) cassettes can negatively affect the lentiviral vector titers. We show that shRNAs, which target the vector genomic RNA, strongly reduced lentiviral vector titers but inhibition of the RNAi pathway via saturation could rescue vector production. The presence of miRNAs in the vector RNA genome (sense orientation) results in a minor titer reduction due to Drosha processing. A major cause for titer reduction of miRNA vectors is due to incompatibility of the cytomegalovirus promoter with the lentiviral vector system. Replacement of this promoter with an inducible promoter resulted in an almost complete restoration of the vector titer. We also showed that antisense poly(A) signal sequences can have a dramatic effect on the vector titer. These results show that not all sequences are compatible with the lentiviral vector system and that care should be taken in the design of lentiviral vectors encoding RNAi inducers.

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Section: Inhibition Of the Rnai Pathway To Increase The Lentiviral Vementioning

confidence: 99%

Section: Dna Constructsmentioning

confidence: 99%

Titers of lentiviral vectors encoding shRNAs and miRNAs are reduced by different mechanisms that require distinct repair strategies

Liu¹,

Vink²,

Westerink³

et al. 2010

RNA

View full text Add to dashboard Cite

show abstract

“…For other viruses, a reduction in IFN signalling during infection results in enhanced virus yields, indicating a limiting role of IFN in virus replication (Young et al, 2003;de Vries et al, 2008). In this study, we analysed the impact of IFN signalling on influenza virus replication in MDCK cells, a cell line approved for influenza vaccine production.…”

Section: Introductionmentioning

confidence: 99%

High yields of influenza A virus in Madin-Darby canine kidney cells are promoted by an insufficient interferon-induced antiviral state

Seitz

Frensing

Höper

et al. 2010

Journal of General Virology

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“…Provision of RNAi-silencing suppressors in trans can indeed improve the production of LV, adenoviral vectors, and especially Sindbis virus vectors. 179 Several RNAi-specific problems may be encountered when viral vectors encode shRNAs or miRNAs. First, high transgene expression levels may impact on viability of the vector-producing cell and the eventual amount of vector that is produced.…”

Section: Vector Production Issuesmentioning

confidence: 99%

Improving miRNA Delivery by Optimizing miRNA Expression Cassettes in Diverse Virus Vectors

Herrera-Carrillo

Liu

Berkhout

2017

Human Gene Therapy Methods

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The RNA interference pathway is an evolutionary conserved post-transcriptional gene regulation mechanism that is exclusively triggered by double-stranded RNA inducers. RNAi-based methods and technologies have facilitated the discovery of many basic science findings and spurred the development of novel RNA therapeutics. Transient induction of RNAi via transfection of synthetic small interfering RNAs can trigger the selective knockdown of a target mRNA. For durable silencing of gene expression, either artificial short hairpin RNA or microRNA encoding transgene constructs were developed. These miRNAs are based on the molecules that induce the natural RNAi pathway in mammals and humans: the endogenously expressed miRNAs. Significant efforts focused on the construction and delivery of miRNA cassettes in order to solve basic biology questions or to design new therapy strategies. Several viral vectors have been developed, which are particularly useful for the delivery of miRNA expression cassettes to specific target cells. Each vector system has its own unique set of distinct properties. Thus, depending on the specific application, a particular vector may be most suitable. This field was previously reviewed for different viral vector systems, and now the recent progress in the field of miRNA-based gene-silencing approaches using lentiviral vectors is reported. The focus is on the unique properties and respective limitations of the available vector systems for miRNA delivery.Keywords: miRNA cassettes, gene therapy, viral vectors, vector tropism INTRODUCTION NON-CODING RNA MOLECULES play an essential role in gene regulation of the cell via a mechanism called RNA interference (RNAi). The RNAi mechanism is evolutionary conserved in eukaryotes and triggers the sequence-specific inhibition or degradation of a single or a unique set of complementary mRNAs. Three small RNA classes have been described to participate in mammalian RNAi mechanisms: microRNAs (miRNAs), 1,2 endogenous small interfering RNAs (endo-siRNAs), 3 and PIWI-associated RNAs (piRNAs). 4 The endo-siRNAs and piRNAs are involved in suppression of transposable elements. The miRNAs are broadly involved in the regulated expression of multiple cellular genes and execute their effect at the posttranscriptional level. 5,6 MiRNA-mediated regulation of gene expression plays a significant role in cell metabolism and cellular developmental and differentiation processes in mammals. More than a thousand human miRNAs have already been identified that are involved in the regulated expression of around 30% of all genes, which is a conservative estimate.7 Because of the miRNA abundance and their important regulatory functions, these molecules have received much attention from the scientific community over the last decade. For instance, efforts have been made to explain the biological function of the natural miRNAs by identifying the target mRNA and the role in cellular physiology. On the other hand, the design of man-made miRNA mimics to impose control over gene expression beca...

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Increased virus replication in mammalian cells by blocking intracellular innate defense responses

Cited by 48 publications

References 51 publications

Titers of lentiviral vectors encoding shRNAs and miRNAs are reduced by different mechanisms that require distinct repair strategies

Titers of lentiviral vectors encoding shRNAs and miRNAs are reduced by different mechanisms that require distinct repair strategies

High yields of influenza A virus in Madin-Darby canine kidney cells are promoted by an insufficient interferon-induced antiviral state

Improving miRNA Delivery by Optimizing miRNA Expression Cassettes in Diverse Virus Vectors

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