Sendai virus cantell strain

Manufactured by Charles River Laboratories

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Sendai virus (Cantell strain) is a laboratory-adapted strain of the Sendai virus, a member of the Paramyxoviridae family. The Sendai virus is an RNA virus that is commonly used as a tool in cell biology research. The Cantell strain is a well-characterized variant of the Sendai virus that is widely utilized in various research applications.

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Sendai virus (13 citations) Sendai virus (SeV; Cantell strain) (7 citations) Cantell strain (2 citations)

25 protocols using sendai virus cantell strain

HCV Subgenomic Replicon Cell Lines

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Huh7 and Huh7.5 [21 (link)], Huh7-HCV K2040 replicon cells that propagate culture-adapted variants of the Con1 HCV (genotype 1B) subgenomic replicon RNA [29 (link)], and the non-neoplastic hepatocyte PH5CH8 cells [30 (link)] were cultured according to standard techniques. Sendai virus strain Cantell was obtained from Charles River Laboratory.

Horner S.M., Wilkins C., Badil S., Iskarpatyoti J, & Gale M J.r. (2015). Proteomic Analysis of Mitochondrial-Associated ER Membranes (MAM) during RNA Virus Infection Reveals Dynamic Changes in Protein and Organelle Trafficking. PLoS ONE, 10(3), e0117963.

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Quantification of Viral RNA in Sendai-Infected Cells

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Cells (5 × 10⁵) were plated in a 6-well plate, and the next day, the cells were infected with Sendai virus (strain Cantell; Charles Rivers Laboratories) at 40 hemagglutinating units (HAU)/ml in medium without serum. After 1 h, the medium was replaced with complete medium, and cells were harvested at the indicated time points. Expression of viral antigen was determined on Western blots using anti-Sendai virus antibody. Total RNA was isolated from infected cells using TRIzol reagent (Invitrogen) or a QIAmp viral RNA kit (Qiagen), and qRT-PCR was performed to quantify the viral RNA copy number as described previously (62 (link)).

Manivannan P., Siddiqui M.A, & Malathi K. (2020). RNase L Amplifies Interferon Signaling by Inducing Protein Kinase R-Mediated Antiviral Stress Granules. Journal of Virology, 94(13), e00205-20.

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Zika and Sendai Virus Infections in Human Keratinocytes

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Zika virus–Dakar (DAK; Zika virus/A.africanus-tc/SEN/1984/41525-DAK) (GenBank accession no. KU955591) was used for the in vitro experiments in human keratinocytes, and Zika virus–French Polynesia (PF; Zika virus strain H/PF/2013) (GenBank accession no. KJ776791.2) was used for in vivo infection studies. Strains were provided by S. Weaver at the University of Texas Medical Branch and H.L. (coauthor of this study), respectively. Sendai virus strain Cantell was obtained from Charles River Laboratory. Zika virus stocks were prepared as described (55 (link)). All viruses were titered by focus-forming assay in Vero cells. Vero cells (CCL-81) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA) and were verified as mycoplasma free by the LookOut Mycoplasma PCR Detection Kit (Sigma-Aldrich, St. Louis, MO, USA). In vitro Zika and Sendai virus infections were performed in serum-free media conditions for 1 hour, after which human keratinocyte medium with indicated treatment conditions was replenished.

Kwock J.T., Handfield C., Suwanpradid J., Hoang P., McFadden M.J., Labagnara K.F., Floyd L., Shannon J., Uppala R., Sarkar M.K., Gudjonsson J.E., Corcoran D.L., Lazear H.M., Sempowski G., Horner S.M, & MacLeod A.S. (2020). IL-27 signaling activates skin cells to induce innate antiviral proteins and protects against Zika virus infection. Science Advances, 6(14), eaay3245.

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Plasmids, Viruses, and Reagents for Signaling Studies

Cited 2 times

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Expression plasmids encoding Flag-, Myc-, or HA-tagged wild-type or mutations of human TBK1, IKKε, IRF3, caRIG-I, MAVS, STING, TAZ, YAP, Lats1, Lats2, TRIF, MyD88, K63-Ub, caALK5, and reporters of TCF, Gli1, 4SBE, NF-κB, 5xUAS, IFNβ_Luc and 5xISRE_Luc have been described previously^{19 (link),69 (link)}. YAP truncations including YAP a.a. 1–170, 1–290, 171–488, 291–488, and the GST-tagged YAP full-length and a.a. 291–488 were generated by PCR-based cloning performed by pfu DNA polymerase from Stratagene. Detailed information will be provided upon request. All coding sequences were verified by DNA sequencing.
GFP and Luciferase double tagged HSV-1 was gifted from Dr. Jiahuai Han (Xiamen University, Xiamen), GFP tagged VSV was gifted from Dr. Zhijian J. Chen (UT Southwestern Medical Center, Dallas), and Sendai virus (Cantell strain) was from Charles River Laboratories. The pharmacological reagents BX795 (Millipore), 2-DG (Sangon Biotech), Doxycycline (Sangon Biotech), and poly(I:C) (Invivogen) were purchased.
Detailed information of all antibodies applied in immunoblotting, immunoprecipitation and immunofluorescence is provided in the attached Supplementary Table 2.

Zhang Q., Meng F., Chen S., Plouffe S.W., Wu S., Liu S., Li X., Zhou R., Wang J., Zhao B., Liu J., Qin J., Zou J., Feng X.H., Guan K.L, & Xu P. (2017). Hippo signaling governs cytosolic nucleic acid sensing through YAP/TAZ-mediated TBK1 blockade. Nature cell biology, 19(4), 362-374.

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Viral Infection Methods: HAV, HCV, DENV, and ZIKV

Cited 2 times

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High-titer HAV (HM175/18f strain) stock was mycoplasma-free and prepared as described before^{34 (link)}. HAV infection was performed at an m.o.i of 10. Sendai virus (Cantell strain) was obtained from Charles River Laboratories and was inoculated at 50 units ml⁻¹, unless otherwise indicated. Infection with HCV carrying Gaussia luciferase (GLuc) reporter was described previously^{32 (link)}. Dengue virus (DENV) serotype 2 (o1Sa-054 strain) and Zika virus (ZIKV, MR-766 and AB-59 strains) were propagated in Vero, C6/36, or Huh-7.5 cells as described^{35 (link)} and inoculated at an m.o.i. of 1.

Yamane D., Feng H., Rivera-Serrano E.E., Selitsky S.R., Hirai-Yuki A., Das A., McKnight K.L., Misumi I., Hensley L., Lovell W., González-López O., Suzuki R., Matsuda M., Nakanishi H., Ohto-Nakanishi T., Hishiki T., Wauthier E., Oikawa T., Morita K., Reid L.M., Sethupathy P., Kohara M., Whitmire J.K, & Lemon S.M. (2019). Basal expression of interferon regulatory factor 1 drives intrinsic hepatocyte resistance to multiple RNA viruses. Nature microbiology, 4(7), 1096-1104.

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MAVS-/- MEF Cell Infection Assay

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MAVS−/− MEF cells as described23 (link) were cultured in DMEM medium supplemented with FBS (fetal bovine serum; 10%, ExCell Bio, FSP500), penicillin (100 U ml⁻¹) and streptomycin (100 μg ml⁻¹). HEK 293T cells were as described23 (link) and were grown in DMEM medium supplemented with 10% calf serum and antibiotics. Recombinant virus VSV-ΔM51-GFP (ref. 42 (link)) was amplified in Vero cells18 (link) and used with a multiplicity of infection=1. For VSV infection, MAVS−/− MEF cells were seeded in 12-well plates at a density of 1 × 10⁵ cells per well. Before incubation with 0.5 ml VSV in DMEM medium MEF cells were washed once with PBS. One hour after incubation, equal volume of DMEM with 20% FBS and antibiotics was added to the cells. Twenty-four hours after infection, cells were harvested for following analysis. Sendai virus (Cantell strain) was from Charles River Laboratories and used at a concentration of 100 HA units ml⁻¹.

Shi Y., Yuan B., Qi N., Zhu W., Su J., Li X., Qi P., Zhang D, & Hou F. (2015). An autoinhibitory mechanism modulates MAVS activity in antiviral innate immune response. Nature Communications, 6, 7811.

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Virus Infection and Analysis in Cells

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HEK293 cells or primary macrophages were infected with Sendai virus (Cantell strain, Charles River Laboratories) at 30–80 HA units for 18 hr, and processed for q-RTPCR. A genetically modified Influenza A virus (IAV) (A/Puerto Rico/8/34) with green fluorescent protein insertion was propagated in MDCK cells and used for the studies [68] (link). The HEK293 cells were infected with IAV at an MOI of 2 for 12 hr, fixed and microscopy was performed.

Pulloor N.K., Nair S., Kostic A.D., Bist P., Weaver J.D., Riley A.M., Tyagi R., Uchil P.D., York J.D., Snyder S.H., García-Sastre A., Potter B.V., Lin R., Shears S.B., Xavier R.J, & Krishnan M.N. (2014). Human Genome-Wide RNAi Screen Identifies an Essential Role for Inositol Pyrophosphates in Type-I Interferon Response. PLoS Pathogens, 10(2), e1003981.

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Rotavirus and Cholera Toxin Protocols

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Rhesus rotavirus (RRV) was provided by Nicholas C. Zachos (Johns Hopkins University, Baltimore, MD, USA). For human enteroids, human rotavirus strain, Ito, was used as previously described (Saxena et al., 2015 (link)). Sendai virus (Cantell strain) was purchased from Charles River Laboratory. B subunit of cholera toxin (CTxB) was obtained from Calbiochem. Monovalent cholera toxin was described previously (Jobling et al., 2012 ). B-subunit of Shiga toxin (STxB) was described previously (Day et al., 2015 (link)).

Maeda K., Zachos N.C., Orzalli M.H., Schmieder S.S., Chang D., Gwilt K.B., Doucet M., Baetz N.W., Lee S., Crawford S.E., Estes M.K., Kagan J.C., Turner J.R, & Lencer W.I. (2022). Depletion of the apical endosome in response to viruses and bacterial toxins provides cell-autonomous host defense at mucosal surfaces. Cell host & microbe, 30(2), 216-231.e5.

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TLR Ligand Activation Assay

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The following ligands were used: Pam3CSK4 (TLR1/2; tlrl-pms; Invivogen, 200 ng/ml), FSL1 (TLR2/6; tlrl-fsl; Invivogen, 100 ng/ml), Poly (I:C) (TLR3; vac-pic; Invivogen, 10 μg /ml), LPS K12 (TLR4; tlrl-eklps; Invivogen, 200 ng/ml), Flagellin (TLR5; tlrl-stfla; Invivogen, 100 ng/ml), R837 (TLR7; tlrl-imqs; Invivogen, 10 μg/ml), R848 (TLR7/8; tlrl-r848; Invivogen, 100 ng/ml), CL075 (TLR8; tlrl-c75; Invivogen, 5 μg/ml), and CpG2006 (TLR9; tlrl-2006; Invivogen, 10 μM), and human interferon-beta 1a (100U/ml) was purchased from PeproTech GmbH (#300–02BC; Germany), 2’3’-cGAMP (InvivoGen, 1ug/ml), Lipofectamine 3000 (Invitrogen), diABZI STING agonist-1 (MedChemExpress, 1μM), Sendai Virus (Cantell Strain) (Charles River, 10 HA units/ml), and HSV-1 (MOI = 1), infected as previously described [28 (link)].

Kim H., Subbannayya Y., Humphries F., Skejsol A., Pinto S.M., Giambelluca M., Espevik T., Fitzgerald K.A, & Kandasamy R.K. (2021). UMP-CMP kinase 2 gene expression in macrophages is dependent on the IRF3-IFNAR signaling axis. PLoS ONE, 16(10), e0258989.

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Sendai Virus Cantell Strain Acquisition

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Sendai Virus-Cantell strain was purchased from Charles River Labs and obtained from Vincent Racaniello (Columbia University).

Flores M., Chew C., Tyan K., Huang W.Q., Salem A, & Clynes R. (2015). FcγRIIB prevents Inflammatory Type I Interferon Production from pDCs during a Viral Memory Response. Journal of immunology (Baltimore, Md. : 1950), 194(9), 4240-4250.

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