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Sindbis Virus

Sindbis virus is a single-stranded RNA virus belonging to the Togaviridae family.
It is an arthropod-borne virus that can infect a variety of vertebrate hosts, including humans.
Sindbis virus is the prototype member of the Alphavirus genus and is known to cause a mild febrile illness in humans, sometimes accompanied by rash and arthralgia.
Researchers studying Sindbis virus can optimize their work using PubCompare.ai's AI-driven platform, which helps locate the best protocols from literature, preprints, and patents using advanced search and comparison tools.
This intuitve interface allows scientists to discover the most effective methods and prodcuts for their Sindbis virus studies.

Most cited protocols related to «Sindbis Virus»

1
Comparative Viral Inactivation Study
Cited 14 times
All flavivirus and non-flavivirus strains listed below were derived from cell culture and provided by the Robert Koch Institute, Berlin, Germany. The following inactivated and stable virus preparations were used in this study: DENV-1 VR344 (Thai 1958); DENV-2 VR345 (TH-36 strain); DENV-3 VR216 (H87 strain); DENV-4 VR217 (H241 strain); WNV Uganda strain (AY532665); WNV Israel (H. Bin, Sheba Medical Center, Israel), kunjin virus (Institute Pasteur, France), usutu virus (AY453411); JEV (ATCC SA14-14-2); Saint Louis encephalitis virus (SLEV) (ATCC VR-1265); TBEV strain K23 (AF091010); YFV strain 17D (X03700); YFV strain ASIBI (AY640589); YFV strain Brazil; YFV strain Ivory Coast; Russian Spring Summer Encephalitis virus (RSSEV); chikungunya virus (LR 2006); chikungunya virus African isolate; sindbis virus; Rift Valley Fever virus and influenza A virus subtype H5N1 (A/dk/Germany R603/06 H5N1).
Two different standard preparations of H1N1 influenza viruses (A/California/04/2009 and A/Hamburg/04/2009) used in this study were provided by the European Network for Diagnostics of Imported Viral Diseases (ENIVD). RNA of Nounane virus was kindly provided by Dr. Fabian Leendertz from NG2, Robert Koch Institute, Berlin. RNA of eleven African flaviviruses (Table 3) were kindly provided by Dr. Amadou A Sall and testing of these viruses was done at Institut Pastuer Dakar, Senegal.
Viral RNA from above described viruses was isolated from 140 μl aliquots of cell culture supernatants, using the QIAamp Viral Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. RNA was eluted in 100 μl of elution buffer and stored at -80°C until further use.
Patel P., Landt O., Kaiser M., Faye O., Koppe T., Lass U., Sall A.A, & Niedrig M. (2013). Development of one-step quantitative reverse transcription PCR for the rapid detection of flaviviruses. Virology Journal, 10, 58.
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Publication 2013
Buffers Cell Culture Techniques Chikungunya virus Diagnosis Encephalitis, Far Eastern Russian Encephalitis Virus, St. Louis Europeans Flavivirus H 241 Influenza A virus Influenza in Birds Kunjin virus Negroid Races Orthomyxovirus Type A, Porcine Rift Valley fever virus RNA, Viral RNA Viruses Sindbis Virus Standard Preparations Strains Thai Usutu virus Virus Virus Diseases
2
Evaluating Cryo-EM Focus Determination Methods
Cited 10 times

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Publication 2012
Bacteriophage T7 Capsid Proteins Sindbis Virus Virion
3
Inducible Dual-Luciferase Assay in Drosophila S2 Cells
Cited 6 times
Drosophila S2 cells (Invitrogen) were cultured at 25°C in Schneider's Drosophila medium (GIBCO), supplemented with 10% heat inactivated foetal calf serum, 2mM L-glutamine, 100 U/ml penicillin, and 100 mg/ml streptomycin. Firefly (Photinus pyralis) and Renilla reniformis luciferase sequences from the plasmids pGL3 and pRL-CMV (Promega) were cloned into pMT/V5-HisB (Invitrogen), generating pMT-Luc and pMT-Ren allowing Copper-inducible expression from a metallothein promoter.
Transfections were performed using Effectene transfection reagent (Qiagen) according to the manufacturer's recommendations. Luciferase expression was assayed using the Dual-Luciferase Reporter Assay System (Promega) and analyzed on a Tecan Ultra-evolution platereader. Double stranded RNA was generated by in vitro transcription from T7 promoter flanked PCR products. Viral stocks were prepared on low passage S2 cells and titered by end point dilution. Briefly, 25.000 S2 cells per well in a 96 well plate were inoculated with 10-fold dilutions of viral stocks. Cells were transferred to fresh medium at day 7 and CPE was monitored visually over 14 days. Viral titers were calculated according to the method of Reed and Muench.
Recombinant Sindbis virus expressing GFP during viral replication was generated by cloning enhanced GFP into the XbaI site of the double subgenomic Sindbis vector pTE3′2J (kindly provided by C. Rice)1 (link). The resulting plasmid was linearized and in vitro transcribed using the mMessage machine kit (Ambion). RNA was purified and electroporated into BHK-21 cells and supernatant was harvested and virus title determined by plaque assay on BHK cells.
Saleh M.C., Tassetto M., van Rij R.P., Goic B., Gausson V., Berry B., Jacquier C., Antoniewski C, & Andino R. (2009). Antiviral immunity in Drosophila requires systemic RNAi spread. Nature, 458(7236), 346-350.
Publication 2009
Biological Assay Biological Evolution Cells Cloning Vectors Copper Dental Plaque Drosophila Effectene Fetal Bovine Serum Fireflies Glutamine Luciferases Luciferases, Renilla Oryza sativa Paragangliomas 3 Penicillins Photinus Plasmids Promega RNA, Double-Stranded Sindbis Virus Streptomycin Technique, Dilution Transcription, Genetic Transfection Virus Virus Replication
4
Sindbis Virus Infection in Honey Bees
Cited 5 times
Sindbis virus expressing enhanced GFP (SINV-GFP) was produced by transfecting in vitro transcribed RNA from the pTE3’2J with GFP inserted into the XbaI site into BHK cells [15 (link),77 (link)]. SINV-GFP was titered on BHK cell monolayers via plaque assay. SINV-GFP (3,750 plaque forming units (PFUs)) was diluted in buffer (2 μl of 10 mM Tris, pH 7.5) and injected into the thorax using a Harbo syringe (from Honey Bee Insemination Service; www.honeybee.breeding.com/HarboAssembly.html) equipped with disposable borosilicate needles made from capillary tubes (0.8-1.10 x 100 m) using a micropipette puller (Sutter Instruments Model P-87). Honey bees were immobilized via incubation in a cold room (4°C) for 20 minutes and with insect pins and forceps during injection; after injection bees recovered at room temperature within 5 minutes. Honey bees were infected with 3,750 PFUs of SINV per bee. This virus dose is modest compared to Drosophila studies which typically utilize 250-2500 PFUs per fly; a newly emerged female worker honey bee (~150 mg) weighs ~ 200x more than an adult female fruit fly (0.8 mg)[15 (link)]. This dose allowed for a natural progression of infection over the course of the experiment and at 3-days post-injection (p.i.) virus abundance could be easily assessed by microscopy and Western-blot analysis. Virus was either injected alone or in conjunction with dsRNA (1 μg).
Flenniken M.L, & Andino R. (2013). Non-Specific dsRNA-Mediated Antiviral Response in the Honey Bee. PLoS ONE, 8(10), e77263.
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Publication 2013
Biological Assay Buffers Capillaries Cells Chest Common Cold Dental Plaque Disease Progression Drosophila Females Forceps Honey Infection Insecta Insemination Microscopy Needles RNA, Double-Stranded Sindbis Virus Syringes Tromethamine Virus Western Blot Woman Workers
5
Dissociated Postnatal Rat Hippocampal Neuron Cultures
Cited 5 times

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Publication 2010
Cells Cloning Vectors Diploid Cell Infection Lysine Neurons Perfusion Poly A Population Group Sindbis Virus Virus

Most recents protocols related to «Sindbis Virus»

1
SARS-CoV-2, Chikungunya, and Other Viral Infections
MRC-5 cells (human lung fibroblasts, ATCC-CCL-171) were cultured essentially as described previously [30 (link)]. Vero E6 cells (African green monkey kidney epithelial cells), 293/ACE2 cells (originally described to be derived from human HEK293 cells [95 (link)], but most likely from nonhuman primate origin [14 (link)]) and BHK-21 cells (baby hamster kidney fibroblasts) were cultured essentially as described previously [21 (link)]. Vero cells (ATCC-CCL81) cells were cultured essentially as described previously [96 (link)]. All culture media contained 100 IU/ml penicillin and 100 ug/ml streptomycin unless otherwise specified.
CHIKV LS3 is a synthetic Chikungunya virus based on the consensus sequence of E1-226V isolates [21 (link)]. Infections with CHIKV LS3 and LS3-GFP were performed essentially as described previously [21 (link)]. The Chikungunya replicon was derived from CHIKV LS3 by replacing the structural genes with a puromycin resistance/foot-and-mouth disease virus 2A oligopeptide/green fluorescent protein (PAC-2A-GFP) reporter gene. The sequence of the 2A oligopeptide contains an amino acid motif that prevents formation of the peptide bond between glycine and the final proline of the sequence which allows expression of multiple proteins from a single ORF [97 (link),98 (link)].
Vero E6 cells were infected with the Sindbis virus (SINV) HR-strain [99 (link)], Semliki Forest virus (SFV) strain SFV4 [100 (link)] or VEEV vaccine strain TC-83 [101 (link)] at a multiplicity of infection (MOI) of 5 or 10. Vero E6 cells were infected with a GFP-expressing recombinant human adenovirus type 5 (HAdV-GFP/LUC; [102 (link)]) at an MOI of 5 and with a GFP-expressing recombinant coxsackie B3 virus (CVB3) (a kind gift from prof. dr. Frank van Kuppeveld, Utrecht University) at an MOI of 1. BHK-21 cells were infected with the equine arteritis virus (EAV) Bucyrus strain at MOI 5 at 39.5°C essentially as described previously [103 (link)].
Treffers E.E., Tas A., Scholte F.E., de Ru A.H., Snijder E.J., van Veelen P.A, & van Hemert M.J. (2023). The alphavirus nonstructural protein 2 NTPase induces a host translational shut-off through phosphorylation of eEF2 via cAMP-PKA-eEF2K signaling. PLOS Pathogens, 19(2), e1011179.
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Publication 2023
ACE2 protein, human Adenoviruses, Human Cells Cercopithecus aethiops Chikungunya Fever Chikungunya virus Consensus Sequence Coxsackievirus Infections Culture Media Epithelial Cells Equine Arteritis Virus Fibroblasts Foot-and-Mouth Disease Virus Genes Genes, Reporter Glycine Hamsters HEK293 Cells Homo sapiens Infant Infection Kidney Lung Oligopeptides Penicillins Peptide Biosynthesis Proline Proteins Puromycin Replicon Reproduction Semliki forest virus Sindbis Virus Strains Streptomycin Vaccines Vero Cells
2
Viral Titer Determination by Plaque Assay
Sendai virus (SeV) Cantell strain was purchased from ATCC (#VR-907). SeV viral titer was determined by plaque assay using LLC-MK2 cells. Cells were plated and infected with SeV in 200 μL of serum-free medium for 1 h for adsorption. Then, LLC-MK2 cells were overlaid with agarose (0.45% in culture media supplemented with 5 µg/mL acetylated trypsin). After 5 days, cells were fixed with trichloroacetic acid (10%) for 30 min, stained with crystal violet (0.1% crystal violet / 25% EtOH) for 5 min, and counted to determine viral titer. Sindbis virus (SINV) Ar-339 strain was purchased from ATCC (#VR-1585). SINV viral titer was determined by plaque assay using Vero cells. Cells were plated into 6 well dished and the adsorption of the virus was performed for 1 h at 37 °C in 200 µL of virus diluted in serum-free DMEM. Then, Vero cells were overlaid with agarose (0.45% in culture media). Poliovirus type 1 (PV) (Mahoney strain) was obtained from Dr. Eckard Wimmer at Stony Brook University, New York. PV viral titer was determined by plaque assay using HeLa cells. Cells were plated into 6 well dished and adsorption of the virus was performed for 30 min at room temperature in 200 µL of virus diluted in serum-free DMEM. Then, cells were overlaid with agarose (0.45% in culture media).
Manjunath L., Oh S., Ortega P., Bouin A., Bournique E., Sanchez A., Martensen P.M., Auerbach A.A., Becker J.T., Seldin M., Harris R.S., Semler B.L, & Buisson R. (2023). APOBEC3B drives PKR-mediated translation shutdown and protects stress granules in response to viral infection. Nature Communications, 14, 820.
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Publication 2023
Adsorption Biological Assay Calculi Cells Culture Media Dental Plaque Ethanol HeLa Cells Human poliovirus 1 Sendai virus Sepharose Serum Sindbis Virus Strains Trichloroacetic Acid Trypsin Vero Cells Violet, Gentian Virus
3
Sindbis Virus Infection in Aedes aegypti
Adult female Ae. aegypti (2–3 days post emergence) were infected with SINV strain MRE-16, and p5’dsMRE16ic-mCh-Flag and p5’dsMRE16ic-AaBec-mCh-Flag by intrathoracic injection. Briefly, each mosquito was cold-anesthetized using a cooling plate and injected with ~10,000 focus-forming units (FFU) of the selected viral strain in a total volume of 100 nL. Microinjected mosquitoes were placed in cardboard cages with ad libitum access to 10% sucrose solution and maintained in the insectary under the same conditions as listed above. After 5 and 7 days post injection (dpi), mosquitoes were sacrificed by overexposing them to triethylamine (Sigma, St. Louis, MO, USA) and placed in 2 mL tubes filled with 1 mL of mosquito diluent (20% heat-inactivated fetal bovine serum (FBS) in Dulbecco’s phosphate-buffered saline (PBS), 50 μg/mL penicillin/streptomycin, 50 μg/mL gentamicin, and 2.5 μg/mL fungizone) and a single zinc-plated steel 4.5-mm bead (Daisy, Rogers, AR, USA). Experimental infections were completed across two replicate batches using the same viral stocks and two subsequent mosquito generations. The results of both replicates were pooled for analysis. A total of 83 mosquitoes were analyzed in the SINV-WT group (46 at 5 dpi and 37 at 7 dpi), 74 mosquitoes in the SINV-mCh-Flag group (46 at 5 dpi and 37 at 7 dpi) and 68 mosquitoes in the SINV-AaBec-mCh-Flag group (26 at 5 dpi and 42 at 7 dpi). Mosquito samples were homogenized at 30 Hz for 2 minutes using a TissueLyser II (QIAGEN GmbH, Hilden, Germany) and centrifuged for 60 s at 11,000 rpm. All samples were further tested for the presence of viral infectious particles by focus forming assay as previously described with one minor modification: viral antigens in infected cells were labeled using Sindbis virus immune ascetic fluid (VR-1248AF; 1:500 dilution). 49 (link) Virus titers were calculated and expressed as FFU/mL. Data were analyzed using GraphPad Prism version 8.4.3. Kruskal Wallis tests were used to compare viral titers across mosquito groups.
Pujhari S., Heu C.C., Brustolin M., Johnson R.M., Kim D, & Rasgon J.L. (2023). Sindbis virus is suppressed in the yellow fever mosquito Aedes aegypti by ATG-6/Beclin-1 mediated activation of autophagy. bioRxiv.
Publication Preprint 2023
Antigens, Viral asparaginic acid alpha-tert-butyl ester copper (II) complex Biological Assay Cells Common Cold Culicidae DNA Replication Fetal Bovine Serum Fungizone Gentamicin Infection Penicillins Phosphates prisma Saline Solution Sindbis Virus Steel Strains Streptomycin Sucrose Technique, Dilution triethylamine Virus Diseases Woman Zinc
4
MAPseq with Modified Sindbis Virus
Modified sindbis virus for MAPseq was obtained from the MAPseq Core Facility at Cold Spring Harbor Laboratory7 (link). The viral library used in this study had a diversity of 20,000,000 unique barcodes. Retro-AAV2 coding for mCherry (pAAV2-hSyn-mCherry, Addgene #114472, 2 × 1013 GC/ml) and green fluorescent protein (pAAV2-hSyn-eGFP, Addgene #50465, 2.2 × 1013 GC/ml) under the human synapsin promoter were obtained from Addgene. All viruses were stored at −80°C and aliquots were thawed over wet ice immediately prior to injection.
Zeisler Z.R., London L., Janssen W.G., Fredericks J.M., Elorette C., Fujimoto A., Zhan H., Russ B.E., Clem R.L., Hof P.R., Stoll F.M, & Rudebeck P.H. (2023). High-throughput sequencing of macaque basolateral amygdala projections reveals dissociable connectional motifs with frontal cortex. bioRxiv.
Publication Preprint 2023
cDNA Library Common Cold Green Fluorescent Proteins Homo sapiens Sindbis Virus Synapsins Virus
5
Sectioning and Dissection of Virus-Injected Brain Tissue
From brains injected with sindbis virus, tissue was sectioned at 200 μm on a Leica 3050S cryostat that had been cleaned with RNAseZap prior to use. Sections were collected over dry ice and stored at −80°C prior to dissection. Cortical areas were then dissected according to sulcal landmarks over dry ice. The areas that were collected and our operational definitions of their boundaries can be found in this table.
Samples from each area were combined across 3 sections in the anterior/posterior plane into 1.5-ml Eppendorf tubes, which were stored at −80°C prior to shipping frozen on dry ice for sequencing.
Zeisler Z.R., London L., Janssen W.G., Fredericks J.M., Elorette C., Fujimoto A., Zhan H., Russ B.E., Clem R.L., Hof P.R., Stoll F.M, & Rudebeck P.H. (2023). High-throughput sequencing of macaque basolateral amygdala projections reveals dissociable connectional motifs with frontal cortex. bioRxiv.
Publication Preprint 2023
Brain Cortex, Cerebral Dissection Dry Ice Freezing Sindbis Virus Tissues

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More about "Sindbis Virus"

Sindbis virus is a single-stranded RNA arbovirus belonging to the Togaviridae family and the Alphavirus genus.
This arthropod-borne virus can infect a variety of vertebrate hosts, including humans, and is known to cause a mild febrile illness sometimes accompanied by rash and joint pain (arthralgia).
Researchers studying Sindbis virus can optimize their work using advanced search and comparison tools like PubCompare.ai's AI-driven platform.
This intuitive interface allows scientists to easily locate the most effective protocols, methods, and products from literature, preprints, and patents for their Sindbis virus research.
Key considerations for Sindbis virus studies may include cell culture techniques using media like Fetal Bovine Serum (FBS), Penicillin-Streptomycin (PSinRep5), and Dulbecco's Modified Eagle Medium (DMEM).
Transfection reagents such as Lipofectamine 2000 can facilitate gene expression studies.
RNA extraction kits like the RNeasy Mini Kit and TRIzol can be used for viral RNA isolation and analysis.
Antibodies like Anti-GluA1-N may be employed for protein detection.
Viral RNA synthesis can be supported by tools like the MEGAscript SP6 kit.
Additionally, supplements such as L-glutamine and Minimum Essential Medium (MEM) may be relevant for cell culture optimization.
By leveraging the latest tools and techniques, researchers can enhance the efficiency and impact of their Sindbis virus investigations.