Journal: bioRxiv

Article Title: Generation of broad-spectrum vaccine platforms against tick-borne flaviviruses using ancestral sequence reconstruction

doi: 10.1101/2025.10.31.685878

Figure Lengend Snippet: (A) Plasmid constructs were designed to function as vaccine platforms for high level expression of ancestral TBF envelope antigen using an alphavirus infectious clone platform (SINV WT). A gBlock consisting of the TBF +E sequence was inserted into a Sindbis virus infectious clone following a 3’ subgenomic promoter (3’ SGP) via restriction enzyme digestion and Gibson Assembly (SINV +E). (B) Viral titers of passage 1 (p1) from plasmid-transfected BHK-21 cells. BHK cells were infected at MOI=0.01. Supernatant was collected at indicated times and viral titers were determined by standard plaque assay. (C) Cycle threshold (Ct) values of p1 SINV WT (red) and SINV +E (blue) stocks probing for the ancestral TBF E (ASR TBF E).

Article Snippet: Briefly, SINV WT plasmid was digested with XbaI restriction enzyme (RE) (New England Biolabs, R0145S) to generate a linear product and gel extracted.

Techniques: Plasmid Preparation, Construct, Expressing, Sequencing, Virus, Transfection, Infection, Plaque Assay

Journal: bioRxiv

Article Title: Generation of broad-spectrum vaccine platforms against tick-borne flaviviruses using ancestral sequence reconstruction

doi: 10.1101/2025.10.31.685878

Figure Lengend Snippet: Overview of animal studies in C57BL/6J mice. Female mice (n=8; 9 weeks old) were vaccinated with 10 5 PFU of SINV +E (blue) subcutaneously (SC) in 50 μL solution at day 0 and day 21. Controls include mice vaccinated with 10 5 PFU SINV WT (mock-vaccinated; red) and PBS (mock challenged; green). Whole blood was harvested and sera isolated by cardiac puncture at day 35 to assess neutralizing antibody titers. At day 35 mice are challenged with a lethal dose of 10 5 PFU of Powassan virus SC diluted in 50 μL solution PBS. Weight, clinical score, and survival were monitored over 21 days post-challenge until end of study. Figure created in https://BioRender.com .

Article Snippet: Briefly, SINV WT plasmid was digested with XbaI restriction enzyme (RE) (New England Biolabs, R0145S) to generate a linear product and gel extracted.

Techniques: Isolation, Virus

Journal: Virus Research

Article Title: Mosquito-borne alphaviruses in Zambia: Isolation and characterization of Eilat and Sindbis viruses

doi: 10.1016/j.virusres.2025.199604

Figure Lengend Snippet: Growth kinetics of Zambian SINV isolate M115 Vero (A) and BHK-21 (B) cells were infected with SINV isolate M115 and AR339 at an MOI of 0.0001. C6/36 cells (C) were infected with SINV at an MOI of 0.01. Titers at the indicated each time point were determined by plaque assay. The values shown are mean ± standard deviation (SD) of triplicate. * p < 0.05, *** p < 0.001 by two-way ANOVA test.

Article Snippet: The SINV AR339 strain (VR-1248) was obtained from the ATCC.

Techniques: Infection, Plaque Assay, Standard Deviation