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How many viruses are needed to form a plaque? - 1 views

www.virology.ws/...es-are-needed-to-form-a-plaque
shared by Casey Finnerty on 07 Sep 16 - No Cached
  • Casey Finnerty on 07 Sep 16
     
    This blog post has a more detailed description of virus kinetics.
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Animal viruses with separately packaged RNA segments - 12 views

www.virology.ws/...parately-packaged-rna-segments
shared by Casey Finnerty on 02 Sep 16 - No Cached
  • only recently have examples of such viruses that infect animals been discovered (paper link).
  • six new viruses with segmented RNA genomes, which was confirmed by gel electrophoresis of RNA extracted from virus particles.
  • results of RNA transfection experiments indicated that this RNA is not needed for viral infectivity.
  • ...1 more annotation...
  • RNA viruses with segmented genomes are common, but in this case, the surprise came when it was found that the dose-response curve of infection for these viruses was not linear. In other words, one virus particle was not sufficient to infect a cell (illustrated).
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A Multicomponent Animal Virus Isolated from Mosquitoes: Cell Host & Microbe - 68 views

www.cell.com/...S1931-3128(16)30310-9
shared by kwedul on 02 Sep 16 - Cached
  • reverse genetics system
    • Casey Finnerty
      Casey Finnerty on 02 Sep 16
       
      Please be ready to explain why reverse genetics was a necessary approach for studying this virus.
  • GCXV
    • Casey Finnerty
      Casey Finnerty on 14 Sep 16
       
      Here is an example of a stickie note.
  • Viral replication was detected in three mosquito cell lines and in intrathoracically inoculated adult female mosquitoes
  • ...14 more annotations...
  • Selective degradation with RNase I demonstrates that GCXV has a single-stranded RNA genome.
    • Casey Finnerty
      Casey Finnerty on 19 Sep 16
       
      RNaseI is specific for ssRNA.
  • reverse genetics system
  • Multicomponent plant viruses were recognized on the basis of deviations from the expected relationship between infectious dose and the number of lesions on infected leaves (i.e., exhibiting multi-hit rather than single-hit kinetics; Flint et al., 2009xPrinciples of Virology Volume I: Molecular Biology. Flint, S.J., Enquist, L.W., Racaniello, V.R., Skalka, A.M., Barnum, D.R., and de Evaluación, E. See all References, Sánchez-Navarro et al., 2013xEffects of the number of genome segments on primary and systemic infections with a multipartite plant RNA virus. Sánchez-Navarro, J.A., Zwart, M.P., and Elena, S.F. J. Virol. 2013; 87: 10805–10815Crossref | PubMedSee all References).
    • jsackett13
      jsackett13 on 19 Sep 16
       
      This was explained in the blog article.
  • of any other group of organisms (
    • aceastep
      aceastep on 19 Sep 16
       
      wow
  • no Illumina reads aligned to the segment 5 sequences from the other isolates, and no contigs from ACH27 exhibited significant similarity to the segment 5 sequences from the other isolates. Therefore, segment 5 appears to be absent from ACH27 and TR7094.
  • However, only four segments were assembled for ACH27 and TR7094 (genome size ∼10.6 kb). For these two isolates, the four assembled segments corresponded to the four largest segments assembled in the other isolates (Figure 1Figure 1, segments 1–4).
  • The phylogeny inferred from segment 5 is also inconsistent with those of the other segments. In addition to the absence of this segment in TR7094 and ACH27, this segment exhibited very low levels of nt divergence (0.4%–2.3%), with most sequence variations only present in a single isolate.
  • with each genome segment separately packaged into virions. Although multicomponent genomes are relatively common among RNA viruses that infect plants and fungi, this method of genome organization has not previously been seen in animal viruses (
    • kwedul
      kwedul on 21 Sep 16
       
      Is this the only thing required for a genome to be considered multicomponent?
  • RNA extracted from purified GCXV particles confirmed the presence of a segmented genome (
    • kwedul
      kwedul on 21 Sep 16
       
      So what exactly is it about this purified GCXV that shows it is segmented?
  • The mosquito pools for both ACH27 and TR7094 contained multiple viruses capable of replicating in mosquito cells, so we were unable to obtain pure cultures for these isolates (
    • kwedul
      kwedul on 21 Sep 16
       
      What is it about there being multiple viruses capable of replicating that made them unable to obtain pure cultures for the isolates?
  • Nuclease digestion assays (Figure 1Figure 1B), along with 5′ and 3′ rapid amplification of cDNA ends (RACE), confirmed that all five genome segments were single-stranded, positive-sense RNA (ssRNA+).
    • kwedul
      kwedul on 21 Sep 16
       
      What about these tests confirms all five genome segments were single-stranded positive-sense RNA?
  • all five segments, (2) segments 1–4, and (3) segments 2–5
    • kwedul
      kwedul on 21 Sep 16
       
      Why did they use these three combinations?
  • the number of plaques decreased more quickly than expected with dilution of the inoculant) (
    • kwedul
      kwedul on 21 Sep 16
       
      Why? What is the significance of this?
  • segments 1–5 (passage 1) and segments 1–4 (passage 2)
    • kwedul
      kwedul on 21 Sep 16
       
      What about a second passage, including 1-4, makes it brighter?
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New Study Finds A Virus That Breaks The Rules Of Viral Infection : Goats and Soda : NPR - 8 views

www.npr.org/...-breaks-the-rules-of-infection
shared by Casey Finnerty on 02 Sep 16 - No Cached
  • I don't think anything else in nature moves this way."
  • Poof! The cell is infected. That's all it takes. One ball, sticking to one cell.
  • "The fifth ball seems to be optional," says Jason Ladner, a genomicist at USAMRIID, who helped discover the virus. Getting the fifth one could control how dangerous the virus is, he says.
  • ...3 more annotations...
  • Then to get infected with the virus, a mosquito needs to catch at least four different colored balls
  • The study is part of a larger project aimed at figuring out what viruses, in addition to Zika and yellow fever, could be lurking inside mosquitoes and possibly waiting to spill over into people.
  • "We're trying to make sure that we're not blindsided when the next virus comes around,"
  • Casey Finnerty on 02 Sep 16
     
    "I don't think anything else in nature moves this way."" ARRRGHGHGHH!
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Quasi-equivalent viruses: a paradigm for protein assemblies - 3 views

www.sciencedirect.com/...S0022283697910688
shared by Casey Finnerty on 07 Sep 16 - No Cached
  • The conclusion suggests that some viral assembly principles are limited paradigms for protein associations occurring in the broader range of cell biology including signal transduction, interaction of transcription factors and protein trafficking.
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Research - A Neuro-Oncology Laboratory at Northwestern University in Chicago - 1 views

www.lesniaklab.com/research.html virotherapy
shared by Casey Finnerty on 12 Oct 16 - No Cached
  • Most adenoviruses that have been historically used for gene therapy have been based on serotype 5 (AdWT). Unfortunately, expression of the primary receptor for Ad5 (the coxsackie-adenovirus receptor, CAR) is highly variable on cancer cells. In fact, several studies have demonstrated a resistance of malignant glioma to adenoviral vectors, a finding that was subsequently attributed to the quantitative deficiency of CAR on brain tumor cells.
  • First, we tested a variety of tumor specific promoters and identified survivin (S) as an excellent tumor specific promoter for transcriptional control of E1a, a gene essential for CRAd replication (J Neurosurg 104:583, 2006; Cancer Biol Ther 6:679, 2007).
  • Based on the above date data, we then created a novel oncolytic adenoviral vector which utilizes the survivin promoter and binds to heparan sulfate proteoglycans expressed on malignant brain tumors and named this new vector CRAd-S-pk7 (Hum Gene Ther 18:589, 2007).
    • Casey Finnerty
      Casey Finnerty on 12 Oct 16
       
      How specific is the binding/tropism? HS is fairly widespread throughout the body.
  • ...6 more annotations...
  • Second, we have identified several receptors that are over-expressed on brain tumor cells and created a series of pseudotyped Ad5 vectors that recognize these receptors
  • Our studies with CRAd-S-pk7 indicate that the virus provides the highest level of viral replication and tumor oncolysis in glioma cell lines. At the same time, we observed minimal viral replication and toxicity in normal human brain. Injection of CRAd-S-pk7 inhibited xenograft brain tumor growth by more than 300%.
  • We were the first group in the country to show that human mesenchymal stem cells can be effectively loaded with a replication competent virus and effectively deliver it to an experimental glioma model
  • When oncolytic vectors are loaded onto stem cells, the virus effectively "hides" from the immune system for an extended period of time. The ability of stem cells to suppress the anti-viral immune response in a permissive and tumor-bearing animal model is the subject of one of our latest manuscript
  • Finally, to further enhance the therapeutic efficacy of stem cells, we have optimized them to specifically traffic to intracranial tumors via genetic modification with single-chain antibodies against antigens expressed on gliomas
  • Our latest work in this area supports the development of neural stem cell based cell carriers for oncolytic virotherapy
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