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.

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.

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.

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.

Biomass conversion into highly useful chemicals
SAPNA JAIN, Alabama State University
This is CURE based course that aims at bridging the gap between theoretical knowledge in chemistry and its practical applications at solving real-world problems. It gives students an opportunity to construct and synthesize their knowledge and skills by learning to apply theoretical knowledge to practice by the laboratory research. The purpose of this course is to acquaint students with the fundamental concepts of chemistry, synthetic methods and techniques. The emphasis will be on novel catalysts synthesis and evaluating their activity towards biomass conversion to liquid fuel and useful chemicals. Students will design synthesize, deduce identities of the biomass conversion products from chemical and spectral clues, and predict reaction products.

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.

Analysis of the effects of protein-protein interactions on signaling through a team-based undergraduate biochemistry laboratory course
Daniela Fera, Swarthmore College
We developed a research-based laboratory course centered on a biological problem involving the B-Raf kinase, specifically the mutant that is commonly found in melanomas. One of the major goals of the project for the students is to generate mutants to determine whether a particular region of the B-Raf protein is critical for the interaction with MEK kinase, a downstream target in the pathway. Students analyze the published B-Raf-MEK crystal structure and choose a mutation to generate in B-Raf or MEK that might alter the dissociation constant (KD) of the complex. They design primers, perform PCR to generate their desired mutant, transform and purify the resulting DNA, express the DNA in E. coli, and purify the protein, all before characterizing it. Characterizing the mutant proteins consist of performing basic pull-downs, western blots, spectroscopic absorbance assays, and biolayer interferometry for binding kinetics. Students also engage in group meeting presentations and journal clubs in which they discuss their work and related primary literature, respectively. Group meeting and journal club discussions provide a forum for students to come up with new ideas to analyze their results, or for future work. Students summarize key results in a final presentation and paper, and develop a research proposal based on their work. Data that students obtain from their mutants provide evidence of the importance of a binding region for B-Raf-MEK complex formation, as well as downstream phosphorylation events. Such data will inform future drug discovery programs, as well as form the foundation for students' work in the course the following year. Because working with mutants can result in unpredictable data and results, students sometimes have to adjust their protocols and repeat experiments. Thus, the CURE format of this course also gives students an opportunity to learn to troubleshoot when things do not work as expected, which helps them learn resiliency in science.

MCC: Malate Dehydrogenase CUREs Community
Ellis Bell, University of San Diego
The Malate Dehydrogenase CUREs Community (MCC) project is designed to facilitate the adoption of effective, protein‐centric, Course Based Undergraduate Research Experiences (CUREs) into teaching labs at a wide variety of undergraduate serving institutions. (Primarily Undergraduate Institutions, Research Intensive Universities and Community Colleges) MCC coordinates and conducts pedagogical research into two major features of CUREs:1) their duration (whole semester versus 5‐6 week modules incorporated into a lab class), and 2) the impact of scientific collaboration between institutions (a key aspect of much modern research). Using validated assessment tools we seek to establish their effects on student confidence, persistence in STEM, and ability to design research experiments and interprete data. To facilitate faculty adoption of CURE approaches the project provides a number of resources. These focus on a variety of research areas related to Malate Dehydrogenase including mechanisms of catalysis and regulation, adaptation and evolution, cofactor specificity, folding and stability and interactions in metabolons. Resources include biologics, experimental protocols and assessment tools. The project also coordinates interactions between courses at different institutions to allow incorporation of scientific collaboration into CUREs. These collaborations also facilitate the use of more sophisticated experimental approaches and broaden the experimental scope of the CUREs.

BASIL (Biochemistry Authentic Scientific Inquiry Laboratory)
Arthur Sikora, Nova Southeastern University; Rebecca Roberts, Ursinus College
This curriculum from the BASIL (Biochemistry Authentic Scientific Inquiry Laboratory) biochemistry consortium aims to get students to transition from thinking like students to thinking like scientists. Students will analyze proteins with known structure but unknown function using computational analyses and wet-lab techniques. BASIL is designed for undergraduate biochemistry lab courses, but can be adapted to first year (or even high school) settings, as well as upper-level undergraduate or graduate coursework. It is targeted to students in biology, biochemistry, chemistry, or related majors. Further details about the BASIL biochemistry consortium can be found on the BASIL blog, http://basiliuse.blogspot.com/ The curriculum is flexible and can be adapted to match the available facilities, the strengths of the instructor and the learning goals of a course and institution. These lessons are often used as part of upper-level laboratory coursework with at least one semester of biochemistry as a pre-requisite or co-requisite. The lab has been designed for classes ranging from 10-24 students (working in teams of two or three) per lab section. This lesson can be adapted to laboratory courses for introductory biology, cell and molecular biology, or advanced biology labs.