Office of Undergraduate Research

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Emphasizing equitable and inclusive access to research opportunities, the University of Maryland's Office of Undergraduate Research (OUR) empowers undergraduates and faculty to engage and succeed in inquiry, creative activity, and scholarship. This collection includes materials shared by undergraduate researchers during OUR events. It also encompasses materials from Undergraduate Research Day 2020, Undergraduate Research Day 2021, and Undergraduate Research Day 2022, which were organized by the Maryland Center for Undergraduate Research.

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Subject: search.filters.subject.virology

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Now showing 1 - 5 of 5
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    Viral Implications of the glyA gene in E. coli
    (2024) Sigalovsky, Nicole; Moronell, Gwen; Artemenko, Allison; Izara, Jamie; Ekbatani, Olivia
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    Defining how two avian double-stranded RNA viruses affect lipid droplet (LD) formation and lipid metabolism in vitro
    (2024) Liu, Ying-Rong (Megan); Kehlbeck, Declan; Egana-Labrin, Sofia; Brodrick, Andrew; Sunny, Nishanth; Broadbent, Andrew; Broadbent, Andrew
    Some mammalian and fish double-stranded (ds)RNA viruses hijack lipid droplets (LDs) during their replication cycle, however, our understanding of how avian dsRNA viruses interact with LDs is incomplete. Here, we report data from avian reovirus (ARV) and infectious bursal disease virus (IBDV)- infected cells. Following ARV infection, there was a significant increase in the number (p < 0.0001) and size (p < 0.0001) of LDs compared to mock-infected controls, 6-24 hours post infection (hpi), in both immortalized LMH cells and primary gut cultures. Treatment with the ACCA inhibitor, TOFA, reduced ARV-mediated LD induction, indicating de novo lipogenesis was partially involved in their formation. Moreover, ARV infection decreased the expression of PGC-1α (p < 0.05) and ELOVL2 (p < 0.001). Previous reports demonstrated that inhibition of PGC-1α and ELOVL expression in mice led to cellular lipid accumulation and hepatic steatosis, suggesting ARV could upregulate LD synthesis by decreasing expression of these genes. However, it is unclear whether LD induction was pro- or anti-viral, since both TOFA treatment to inhibit LD synthesis, and oleic acid/palmitic acid treatment to overexpress LDs did not significantly alter ARV replication. In contrast, infection with IBDV did not substantially increase the size or number of LDs, or significantly reduce the expression of PGC-1α and ELOVL2. In contrast to some other dsRNA viruses like rotavirus and mammalian orthoreovirus, neither ARV nor IBDV cytoplasmic puncta colocalized with LDs. Taken together, our findings demonstrate that the two dsRNA viruses had distinct effects on cellular lipid metabolism.
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    Using Synthetic HSV to Study Viral Pathogenesis and Develop Cancer Therapeutics
    (2024) Duggal, Aryaan; Gonzalez-Juarbe, Norberto
    Background: Herpes Simplex Virus 1 (HSV-1) is a linear dsDNA virus typically associated with ulcer development in the mouth and genital mucous membranes. Often overlooked, is the versatile DNA structure that enables HSV-1 to serve as a platform to study viral pathogenesis. HSV-1 also possesses an inherent oncolytic quality, allowing it to function as a selective therapeutic that can be programmed to target malignant cells while inducing a strong host immune response. Methods: Various endonucleases were used to isolate the genome into 11 smaller fragments. To study the effects on viral pathogenicity and oncolytic properties, we synthesized and inserted novel sequences, with a YCpBAC sequence to allow for selection and transformation into yeast spheroplasts for TAR cloning in vitro. Fragments containing Casp2 coding regions were also tested for viability, by running quantification assays and infection tests on cell lineages. Results: Isolated band size was correlated with theoretical values, indicating accurate construction of all HSV-1 fragments. After screening on -His and -Ura plates, a 200µL aliquot of 40ng/µL TAR2 sample was determined to yield a satisfactory 85% success rate in yeast assembly. The ideal quantity for viral replication in Vero E6 cells was determined to be around 1.0×10^7 cells/cm^2 providing 2.5ml of 100x viral solution. Conclusions: Successful reassembly of HSV-1 demonstrates the feasibility of using HSV-1 as a therapeutic vehicle, useful in treating cancers and understanding neurological diseases. These findings provide a strong foundation for subsequent modifications of HSV-1, which will focus on its effects on a variety of cancer cell lines.
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    Selection of FANA Aptamers to Inhibit Sars-CoV-2 Infection
    (2024) Scheirey, Grace; Meda, Anju; Yang, Grant; DeStefano, Jeffrey; DeStefano, Jeffrey
    Aptamers are short nucleic acid sequences that bind to a particular protein target. Recently, FANA aptamers have been found to be ideal subjects for therapeutic treatments of SARS-CoV-2 as they are highly specific, resistant to degradation, and have low immunogenicity. Previous research has identified a FANA aptamer that binds to the receptor binding domain (RBD) of the SARS-CoV-2 Wuhan strain, blocking infection in an airway epithelium model. However, past research has also revealed the possibility of an aptamer that binds to the S1 subunit of SARS-CoV-2 and blocks infection, even without binding directly to the RBD. Thus, our research is focused on identifying FANA aptamers that bind tightly to the SARS-CoV-2 XBB.1.5 variant’s S1 subunit, with the potential to block viral infection. We identified these aptamers by following the systematic evolution of ligands by exponential enrichment (SELEX) protocol, which involves synthesizing FANA start material, isolating sequences that bind to the protein target using magnetic bead assays, performing PCR reactions, and repeating selection cycles until we have selected for aptamers with tight binding affinity. After 7 selection cycles, we identified 32 potential aptamer sequences using Sanger Sequencing and compared their properties. We found that FANA-R7-17 bound very tightly to XBB.1.5-S1 with a Kd of 46 ± 15 pM, and had a half-life of about 21 minutes. Among all the aptamers we compared, FANA-R7-17 presents the greatest potential as an aptamer to the SARS-CoV-2 XBB.1.5 variant, though future research would need to be conducted to determine its ability to block infection in a human airway epithelium model.
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    Engineering HeLa Cells to Better Support Rhinovirus C Infection In Vitro
    (2024) Swaminathan, Divya; Goldstein, Monty; Pankow, Alec; Scull, Margaret; Scull, Margaret
    Human rhinoviruses (RVs) are non-enveloped, positive-sense, single-stranded RNA viruses that target the respiratory tract for infection, leading to common cold-like symptoms, bronchiolitis, or, in some cases, pneumonia. RVs are classified into three phylogenetically distinct groups: RV-A, RV-B, and RV-C, with RV-C being identified most recently, in 2006. However, while RV-A and RV-B viruses have been studied for decades in HeLa cells, RV-C does not naturally infect conventional cell lines and thus, it has been difficult to propagate and study in the lab. Further, no assay currently exists to quantify infectious RV-C particles, such as a plaque assay or limiting dilution assay. It has been demonstrated that cadherin-related family member 3 (CDHR3) is a critical cellular entry receptor for RV-C, and further, that RV-C may depend on the stimulator of interferon genes (STING) for viral genome replication. To investigate the roles of these proteins in RV-C infectivity, inducible human STING (hSTING) was transfected into HeLa cells that stably express CDHR3. Elevated STING expression was then confirmed in these cells after induction with Doxycycline (Dox). Ongoing studies seek to further explore how the expression of additional ciliated cell-specific host factors - such as Prominin-1 (PROM1) - in HeLa cells impact virus infection in this cell line. The hypothesis is that the combined expression of CDHR3, PROM1, and STING in HeLa cells will enhance entry and replication, facilitating amplification of RV-C in the laboratory and aiding detection of infection via antibody-mediated staining or enhanced cytopathic effects. These engineered HeLa cells would thereby constitute a novel cell line that is more amenable to the establishment of a plaque assay or limiting dilution assay to quantify infectious virus.