2019 Summer Scholars

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Group photo outside on steps


Tatyana Dunn

Tatyana Dunn

North Carolina Central University
Mentor: Ju-ahng Lee
Major: Biology/Pre Med

Project: Hands-on training of modern genetic engineering technologies: Transposon-Mediated transgenesis and targeted genome editing by CRISPR/Cas9

I’m working to understand modern engineering technologies and the use of targeted genome editing by CRISPR/Cas9 to knock out zebrafish tyrosinase (tyr) and knock in lgals3bpb. I have been conducting hands-on experiments to knock in the lgals3bpb gene into the zebrafish genome. I have recently learned to do restriction enzyme mapping, mini and midi kits to extract plasmid DNA, Gel electrophoresis, and injections to run trials to see if the knock in is becoming more efficient through different techniques. Thus far, I have found through various readings and trial runs that CRISPR/Cas9 is efficient at knocking out genes, but knock in efficiency is very low. Out of 300 fish injected, only 3 - 6 survived from each sample injected, but this may have been due to toxicity in the injection mixes. Recently, after extracting plasmid DNA and making new injection mixes, we have had most of our new injected fish survive and will soon be able to tell if the knock in expression is shown. My next step is to identify other methods that can be used to raise the knock-in efficiency of CRISPR/Cas9 in zebrafish.


Natalie Ezem

Natalie Ezem

Duke University
Mentor: Gregory Cole
Major: Sociology/Global Health

Project: Expression pattern of cannabinoid receptors 1 and 2 in zebrafish embryos

I am investigating how 4% Paraformaldehyde (PFA) in Phosphate-Buffered Saline (PBS) impacts cannabinoid receptor genes, also known as cnr1 and cnr2, in zebrafish embryos at one to three days post fertilization. I will be determining the correct cnr1 and cnr2 constructs, synthesizing a cnr1 and cnr2 anti-sense DIG-labelled RNA probe, and performing whole-mount in-situ hybridization on the receptors. Thus far, I have found the cnr2 construct and I am currently redesigning new primers to detect cnr1. Afterwards, I will synthesize probes for these sequences and perform the whole-mount in-situ hybridization.


Dezmond Garrett

Dezmond Garrett

University of North Carolina - Greensboro
Mentor: Hiro Matsunami
Major: Kinesiology

Project: Testing olfactory receptors

The goal of my project is to continue to find new agonist ligands of OR1A1 and OR2W1 by exposing them to various odorants. I am testing linkage of odorants predicted by a computational study of results of the project’s Australian collaborators. I will be using the odorants of their predicted study. The activity and linkage will be monitored by the use of luciferase assay. By using human embryonic kidney cells, we will transfect plasmid of the ORs, luciferase to provide luminescence, M3, the transporter protein RTP1S, SV40, and CRE (which act as promoters of Luciferase) into the cells. Once the cells are transfected with the plasmids, we will inject the odorants into the cells and monitor the luminescence to determine the activation of the receptors by the odorants.


Jonathan Harpe

Jonathan Harpe

North Carolina Central University
Mentor: Tim Veldman
Major: Biology Pre Med/Psychology

Project: Confirming peripheral miRNA biomarkers by RT-qPCR in epilepsy patients

The purpose of this pilot study is to evaluate peripheral blood of epilepsy patients for expression profiles that distinguish epilepsy from healthy individuals and patients with neurological disorders other than epilepsy (e.g., migraines). We hypothesize that molecular signals known as microRNAs are detectable in blood and are associated with epilepsy along with standard clinical assessments. These type of RNA are small noncoding molecules that are ~22 nucleotides in length and function as post-transcriptional inhibitors for protein synthesis. Past studies show that because of the nature of these molecules, they could potentially indicate signs of being biomarkers for diseases that have very poor prognosis and prevention. I am inducting managing patients into the study and collecting blood samples to ultimately answer the question of whether these miRNAs collected from plasma samples are linked to epilepsy. After sample collection, we are expecting to find statistically significant up or down regulated miRNAs from the epilepsy subgroup to be analyze by their properties and structure to find which miRNAs may become potential biomarkers for their disorder.


Hana Lee

Hana Lee

North Carolina Central University
Mentor: Ashley Chi
Major: Biology/Pre Med

Project: The role of MESH1 in an in vivo model of ferroptosis

To test the role of MESH1 in the in vivo ferroptosis models, we knock down the expression of enzyme MESH1 by the tail vein injections of siRNAs that target MESH1 (siMESH1) or non-targeting control (siNT: negative control). As a positive control, we injected liproxstatin that is expected to inhibit ferroptosis, or programmed cell death. We fed these mice with the MCDE diet that has been discovered to induce ferroptosis in the liver. We expected to see a decrease in ferroptosis but an increase in fibrosis markers in the liproxstatin and the siMESH1 groups, compared to the siNT group. Now we are validating the presence of other markers for fibrosis using western blot, qPCR and IF staining. We are testing for markers of ferroptosis and fibrosis using the same method.


Kennedy Lofton

Kennedy Lofton

North Carolina Central University
Mentor: Ornit Chiba-Falek
Major: Pharmaceutical Sciences

Project: Using the methylation of DNA to explain a track for gene therapy of AD patients

APOE is correlated with a person having a higher risk of developing Alzheimer’s disease (AD). Therefore, we are investigating if the methylation of AD patients’ DNA changes how their APOE gene is expressed. We are using FANS (Fluorescence-Activated Nuclei Sorting) methodology to sort the cells between neuronal and non-neuronal (glial cells). Next, we are using three different downstream analytical methods to pull different information from the different cell types. For methylation, a MethylEPIC microarray looks at the methylation levels on the cytosine-phosphate-guanine (CpG )islands of the gene. We are also using pyrosequencing to analyze parts of the APOE gene that may lie outside of the CpG islands and their cytosine methylation levels. Finally, we compare data across the different cell types (neuronal and glial), between normal and AD patients, as well as between male and female patients.


Ednan Ochieng

Ednan Ochieng

North Carolina Agricultural and Technical State University
Mentor: Raluca Gordân
Major: Mechanical Engineering

Project: Modeling transcription factor specificity

Using experimental assays to measure the DNA binding specificity of transcription factors (TFs) across the human genome is a long and expensive process. To solve this problem, we are training machine-learning models to predict TF binding specificity. We do this by using a Support-Vector Regression (SVR) algorithm that takes as input TF binding measurements for ~30,000 genomic sites, looks for patterns in the sequences, and returns a function that best predicts the TF specificities from sequence. Longer-term, being able to accurately predict TF binding will allows us to examine gene expression regulation in human cells.


Alexandria Scott

Alexandria Scott

North Carolina Central University
Mentor: Paul Magwene
Major: Biomedical Science

Project: Analyzing capsule size variation in Cryptococcus

Cryptococcus is a pathogenic fungus that primarily infects immune compromised people. To investigate what loci controls the capsule size variation, I will be using natural variation within the C. denoeoformans species to understand the genotype to phenotype relationship regarding virulence factors. Specifically, I will be studying the capsule, which surrounds the cell and protects the fungus from various host specific conditions. A cross was made between two genetically diverse strains, one isolated from pigeon excrement and the other laboratory constructed, and I will use the progeny to study capsule size variation. We will focus on what genes control the capsule size. Due to recombination from the cross, the segregate will have genes from both parents which will be examined further using Quantitative Trait Locus (QTL) analysis of the capsule size variation. The results of QTL analysis will ultimately quantify the genetic differences, some of which are responsible for capsule size.