An Arabidopsis Mutant Screen CURE for a Cell and Molecular Biology Laboratory Course
Jinjie Liu, Michigan State University
This CURE is designed from a crucial component of a chloroplast lipid signaling research project and has been implemented for a cell and molecular biology laboratory course at Michigan State University. The research laboratory generated an engineered plant line producing a lipid-derived plant hormone and mutagenized this line. The research question is "what transporters or receptors are involved in the hormone signaling transduction or perception processes?". Students form research hypotheses based on the research model, design experiments, perform experiments, collect and analyze data, make scientific arguments, and share their findings with the learning community. Specifically, the students culture the mutagenized plant population and select the desired mutant phenotypes, followed by genotyping the mutants and characterizing the mutants by basic biochemical approaches. Mathematics is also integrated into the course design. As the students studied the relevant genetic, molecular and biochemical concepts during this CURE, they use the core idea of information flow and data they generate in the lab to make claims about their mutant plants and support these claims with evidence and reasoning.

Characterizing the Aging Process Using Caenorhabditis elegans and Reverse Genetics
Joslyn Mills, Bridgewater State University
Using gene silencing (RNAi) in the nemotode C. elegans, students will identify genetic modifiers of proteins with roles in aging by reverse genetics. Specifically, students will analyze the effect of knocking down genes on the level of aging-related proteins tagged with fluorophores (GFP, RFP, etc.). Each group of students will use function-specific RNAi libraries (transcription factors, kinases, etc) already established in our lab. Furthermore, students will evaluate the effect of genetic modifiers on proteostasis and lifespan. In addition to becoming familiar with C. elegans work and appreciating the use of model organisms, the students will master microscopy, genetic crosses, gene silencing, and molecular and biochemical readout assays such as qPCR and immunoblotting.

Neurogenetics Laboratory: Mapping a functional circuit for cold nociception in Drosophila
Sarah Clark, Georgia State University
Students will work in small groups to identify neural populations that may be involved in the Drosophila larval response to noxious cold. They will use the GAL4/UAS expression system to excite or inhibit neural populations and assess the impact of their manipulation on the larvae's behavioral response to cold. If a relevant neural population is identified, students will then identify (based on current literature) genes that are likely to be involved in neurite development and/or maintenance in that population. They will use mutations and/or RNA interference to disrupt the function of these genes in the population of interest and assess the effect of the disruption on neuronal morphology and larval behavior.

Exploring eukaryotic protein structure and post-translational modifications.
Erica Jacobs, St. John's University-New York
This CURE will provide opportunity for students to think and act as researchers by using computational, biochemical, and bioanalytical techniques to examine tick antigen proteins. The CURE is designed as a lab for upper-level students who are taking or have taken a one-semester introductory biochemistry course, but two semesters would be even better. It could also be adapted for cell/molecular biology or (bio) analytical chemistry instrumentational analysis labs. It has been taught for classes ranging from 12-24 students. Ticks are notorious vectors of viral, protozoan, and bacterial diseases, including Lyme disease. While an anti-vector vaccine capable of protecting people from diseases transmitted by a particular tick species is an alluring goal, only one such anti-tick vaccine is currently available. This vaccine targets Bm86, a protein from the midgut of Rhipicephalus microplus, a cattle tick. Not only does the vaccine limit parasitism of the cattle by ticks, data suggests that it can also prevent transmission of tick-borne diseases including bovine anaplasmosis and babesiosis. However, similar vaccination approaches have not succeeded thus far against ticks that transmit diseases to humans, and little is known about the antibody response to the antigen, or about the protein itself. Since the protein's structure and function are unknown, the research goal of this CURE is to purify Bm86 using an insect cell/baculovirus expression system and characterize it, including domain structure and post-translational modifications (glycosylation sites). There are homologs to Bm86 in every sequenced tick species examined, and future CUREs will characterize some of the homologs including those in Ixodes scapularis, the tick that is mainly responsible for transmitting Lyme in the eastern US, and Haemaphysalis longicornis, the Asian longhorned tick, a newly-discovered invasive species in the area that also has significant disease-transmitting potential. By understanding the structure and post-translational modifications of this protein, we hope to gain a better understanding of how to make effective anti-tick vaccines, including those for humans, that may prevent transmission of Lyme disease. Importantly, the basic parameters of this CURE can be used to examine other proteins besides tick antigens. For example, during the pandemic, the CURE pivoted from the tick antigen to the SARS-CoV-2 nucleocapsid protein, which was also expressed in an insect cell system. Instead of characterizing glycosylation sites, we characterized phosphorylation sites. It's therefore possible to use this same framework for many different eukaryotic proteins that may be of research interest.

Biochemistry II lab: Crithidia parasite metabolism
Amy Greene, Albright College
Course Undergraduate Research Experiences (CUREs) have been shown to increase student engagement, skills, and retention in STEM. We developed a CURE using non-pathogenic Crithidia fasciculata parasites, which are insect trypanosomes related to the causative agents of Leishmaniasis, African Trypanosomiasis, and Chagas' diseases. This parasite is ideal for undergraduate CUREs because it grows to high density in serum-free inexpensive media, and has not been well studied in the literature, providing opportunities for novel discoveries. Metabolically labelled 1-13C-glucose was added to the parasites, and changes in peak position was monitored over time (either in real time, or in the supernatant). The main fermentation products observed were ethanol and succinate. Student groups then designed a novel project investigating metabolism in Crithidia. Students produced novel data on metabolism in a little-studied parasite.

Population & Community Ecology
Cascade Sorte, University of California-Irvine
Students in a Population and Community Ecology class participate in coastal marine research focused on understanding factors determining population sizes and community interactions, particularly in the context of species that appear to be shifting their ranges with climate change. Students participate in all aspects of the research from making observations and collecting data in the field to defining questions, stating hypothesis, designing and completing statistical analysis, and interpreting and presenting results. The outcomes are a research proposal, research paper, and poster presentation. All are intended to be at a level appropriate for use as a writing sample or presentation at undergraduate conferences. Results are incorporated into the ongoing research project led by the course instructor and graduate student teaching assistant.

Molecular Parasitology
Swati Agrawal, University of Mary Washington
In Spring 2021, we piloted a mini-CURE where student groups from University of Mary Washington and Georgia State University collaboratively completed research projects as part of a research-intensive course on Molecular Parasitology. The benefits of this approach were immediately obvious as students interacted across institutions, learned from each other's disciplinary expertise while informing their own research with data collected by their collaborators.

The HICA project
In this CURE, inspired by the work of Hoffmann, et al., students prepare mutant Haemophilus influenzae carbonic anhydrase (HICA) proteins. Using PyMOL to visualize the three-dimensional structure of the HICA protein, students choose one or more surface amino acid residues to mutate to histidine residues in order to create a surface histidine cluster that will allow the mutant protein to bind to a nickel affinity column. Using site-directed mutagenesis, recombinant plasmids are constructed and are then used to transform an E. coli expression vector. The mutant HICA protein is overexpressed, cells are lysed, and students load the cell lysate onto Ni-NTA columns and determine the imidazole concentration required to elute the mutant protein. The construction of a library of mutant proteins will allow the development of a general method in which specific surface histidine residues of any protein can be mutated in order to facilitate affinity purification. The Haemophilus influenzae bacterium described herein is a respiratory pathogen that causes meningitis (in its encapsulated form) and mucosal infections such as otitis media, sinusitis and conjunctivitis (in its unencapsulated form). A recent study showed that the carbonic anhydrase enzyme is absolutely required for pathogenesis. Furthermore, expression of the HICA enzyme allows the pathogen to survive in host immune cells (Langereis, et al.). These observations make the study of HICA itself particularly attractive, in addition to the overall goal of contributing to a body of work that will allow the minimal histidine character required for nickel affinity to be ascertained.

Isolation and characterization of antibiotic-producing soil bacteria
Maria Messner, Lenoir Community College
One of the biggest threat in hospitals is the rising cases of people who harbor antibiotic-resistant bacterial strains. Therefore, it is critical to find and characterize novel antibiotics to combat the resistant strains. Most of the antibiotics used in healthcare settings come from anti-biotic producing bacteria and fungi found in the soil. The goal of this CURE will be to isolate antibiotic-producing bacteria and fungi from the soil in the local area, and to determine the chemistry of the antibiotics. An extension of the project will be to determine how the presence of antibiotic-producing microbes affect other organisms resident in the soil, as it is unclear as to why microbes use energy to produce antibiotic factors.

Community Flood Risk Assessment from Rising/Surging Seas Project
Kevin Kupietz, Elizabeth City State University
Globally 634 million people, 10% of the world's population, live in coastal areas less than 10 meters above sea level. According to 2010 census data, 123 million people, 39% of the United States population, live in coastal counties with an estimated increase to this number by 8% in the 2020 census. As natural disasters have been seen to increase in frequency and severity in the past five years coupled with expected sea rises from climate change it is important that anyone involved with the safety and resiliency planning of their organization/community have an understanding of how to scientifically assess risk from flooding in order to mitigate and recover from the effects. This project allows students the ability to develop skills to utilize computer modeling systems and to apply the data to real world communities in examining risk to structures as well as different groups in the community.