Short-hairpin RNAs delivered by lentiviral vector transduction trigger RIG-I-mediated IFN activation

Kenworthy, Rachael; Lambert, Diana; Wang, Nan; Chen, Zihong; Zhu, Haizhen; Zhu, Fanxiu; Liu, Chen; Li, Kui; Tang, Hengli

doi:10.1093/nar/gkp714

Cited by 37 publications

(31 citation statements)

References 60 publications

(78 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Specifically, short hairpin RNAs (shRNAs) activate cells via RIG-I and not via TLR3 (Kenworthy et al, 2009), making shRNA an attractive tool for selective ablation of genes of interest in CD8a + cDCs.…”

Section: Discussionmentioning

confidence: 99%

Quantitative Proteomics Reveals Subset-Specific Viral Recognition in Dendritic Cells

et al. 2010

View full text Add to dashboard Cite

Dendritic cell (DC) populations consist of multiple subsets that are essential orchestrators of the immune system. Technological limitations have so far prevented systems-wide accurate proteome comparison of rare cell populations in vivo. Here, we used high-resolution mass spectrometry-based proteomics, combined with label-free quantitation algorithms, to determine the proteome of mouse splenic conventional and plasmacytoid DC subsets to a depth of 5,780 and 6,664 proteins, respectively. We found mutually exclusive expression of pattern recognition pathways not previously known to be different among conventional DC subsets. Our experiments assigned key viral recognition functions to be exclusively expressed in CD4(+) and double-negative DCs. The CD8alpha(+) DCs largely lack the receptors required to sense certain viruses in the cytoplasm. By avoiding activation via cytoplasmic receptors, including retinoic acid-inducible gene I, CD8alpha(+) DCs likely gain a window of opportunity to process and present viral antigens before activation-induced shutdown of antigen presentation pathways occurs.

show abstract

Section: Discussionmentioning

confidence: 99%

Quantitative Proteomics Reveals Subset-Specific Viral Recognition in Dendritic Cells

et al. 2010

View full text Add to dashboard Cite

show abstract

“…Therefore, we concluded that there is no clear difference between genome replications of Ni-CE strain and CE(NiN) strain. On the other hand, it has recently been shown that RIG-I is activated by short double-stranded RNA (19) and short-hairpin RNA (22). Thus, the exact structure of RNA activating RIG-I remains controversial.…”

Section: Discussionmentioning

confidence: 99%

Rabies Virus Nucleoprotein Functions To Evade Activation of the RIG-I-Mediated Antiviral Response

Masatani

Ito

Shimizu

et al. 2010

J Virol

View full text Add to dashboard Cite

The rabies virus Ni-CE strain causes nonlethal infection in adult mice after intracerebral inoculation, whereas the parental Nishigahara (Ni) strain kills mice. We previously reported that the chimeric CE(NiN) strain with the N gene from the Ni strain in the genetic background of the Ni-CE strain kills adult mice, indicating that the N gene is related to the different pathogenicities of Ni and Ni-CE strains. In the present study, to obtain an insight into the mechanism by which the N gene determines viral pathogenicity, we compared the effects of Ni, Ni-CE, and CE(NiN) infections on host gene expressions using a human neuroblastoma cell line. Microarray analysis of these infected cells revealed that the expression levels of particular genes in Ni-and CE(NiN)-infected cells, including beta interferon (IFN-␤) and chemokine genes (i.e., CXCL10 and CCL5) were lower than those in Ni-CE-infected cells. We also demonstrated that Ni-CE infection activated the interferon regulatory factor 3 (IRF-3)-dependent IFN-␤ promoter and induced IRF-3 nuclear translocation more efficiently than did Ni or CE(NiN) infection. Furthermore, we showed that Ni-CE infection, but not Ni or CE(NiN) infection, strongly activates the IRF-3 pathway through activation of RIG-I, which is known as a cellular sensor of virus infection. These findings indicate that the N protein of rabies virus (Ni strain) has a function to evade the activation of RIG-I. To our knowledge, this is the first report that the Mononegavirales N protein functions to evade induction of host IFN and chemokines.Rabies virus, which belongs to Lyssavirus of the family Rhabdoviridae, which belongs to the order Mononegavirales, is known as a highly neurotropic virus and causes fatal encephalitis accompanied by severe neurological symptoms in almost all mammals, including humans. The genome is an unsegmented negative sense RNA and contains five genes (N, P, M, G, and L genes) encoding nucleoprotein (N protein), phosphoprotein (P protein), matrix (M) protein, glycoprotein (G protein), and large (L) protein, respectively (12). The N, P, and L proteins form helical ribonucleoprotein (RNP), together with the viral genomic RNA. The N protein participates in encapsidation of the genomic RNA. Only the encapsidated genomic RNA can be a template for replication of the viral genome and transcription of the viral mRNAs by the RNAdependent RNA polymerase, L protein. The P protein binds to both N and L proteins and functions as a cofactor of the viral RNA polymerase. During virus assembly, the RNP is wrapped into an envelope containing an inner layer of the M protein and the transmembrane spike protein, G protein.In response to viral infection (e.g., picornavirus, bunyavirus, and flavivirus infections), neurons in the brain produce type I interferon (IFN) comprised of the IFN-␣ family and IFN-␤, which induces an antiviral status of a cell and functions as a main player for the host innate immunity (8, 9). The brain neurons are also capable of responding to the produced type I

show abstract

“…More recently, short-hairpin RNAs (shRNAs) delivered by lentiviral vector transduction were shown to trigger RIG-I-mediated IFN activation in a sequence-and 5′-triphosphate-dependent manner. 44 Thus, the shRNA-lentiviral vector also results in undesirable immunostimulation, although viral vector-mediated siRNA/shRNA delivery might be essentially inadequate for clinical use.…”

Section: Figure 4 Isrna With Invivofectamine But Not With Atelocollagmentioning

confidence: 99%

Atelocollagen-mediated Systemic Delivery Prevents Immunostimulatory Adverse Effects of siRNA in Mammals

et al. 2012

View full text Add to dashboard Cite

Short interfering RNA (siRNA) is a potent activator of the mammalian innate immune system. When considering possible clinical applications of siRNA for humans, the adverse immunostimulatory effects must also be taken into account. Here, we show that atelocollagen-mediated systemic delivery of siRNA without chemical modifications did not cause any immunostimulation in both animals and human peripheral blood mononuclear cells (PBMCs), even if the siRNA harbored an interferon (IFN)-inducible sequence. In contrast, systemic delivery of immunostimulatory RNA (isRNA)-mediated by a cationic lipid (such as Invivofectamine) induced potent type-I IFNs and inflammatory cytokines. Regarding the mechanism by which the isRNA/atelocollagen complex avoided adverse effects on immunostimulation, we revealed that this complex was not incorporated into PBMCs. On the other hand, Invivofectamine delivered isRNA into PBMCs. The use of either atelocollagen or Invivofectamine as a vehicle elicited significant and undistinguishable therapeutic effects in a contact hypersensitivity (CHS) inflammatory model mouse, when we intravenously injected the siRNA targeting monocyte chemoattractant protein-1 as the complex. For the goal of realizing siRNA-based medicines for humans, atelocollagen is an excellent and promising delivery vehicle, and it has the useful advantage of evading detection by the "radar" of innate immunity.

show abstract

Short-hairpin RNAs delivered by lentiviral vector transduction trigger RIG-I-mediated IFN activation

Cited by 37 publications

References 60 publications

Quantitative Proteomics Reveals Subset-Specific Viral Recognition in Dendritic Cells

Quantitative Proteomics Reveals Subset-Specific Viral Recognition in Dendritic Cells

Rabies Virus Nucleoprotein Functions To Evade Activation of the RIG-I-Mediated Antiviral Response

Atelocollagen-mediated Systemic Delivery Prevents Immunostimulatory Adverse Effects of siRNA in Mammals

Contact Info

Product

Resources

About