At-Home, Cell-Free Synthetic Biology Education Modules for Transcriptional Regulation and Environmental Water Quality Monitoring

Abstract

As the field of synthetic biology expands, the need to grow and train science, technology, engineering, and math (STEM) practitioners is essential. However, the lack of access to hands-on demonstrations has led to inequalities of opportunity and practice. In addition, there is a gap in providing content that enables students to make their own bioengineered systems. To address these challenges, we develop four shelf-stable cell-free biosensing educational modules that work by simply adding water and DNA to freeze-dried crude extracts of non-pathogenic Escherichia coli. We introduce activities and supporting curricula to teach the structure and function of the lac operon, dose-responsive behavior, considerations for biosensor outputs, and a “build-your-own” activity for monitoring environmental contaminants in water. We piloted these modules with K-12 teachers and 130 high-school students in their classroomsand at homewithout professional laboratory equipment. This work promises to catalyze access to interactive synthetic biology education opportunities.

Ribosome Pool Engineering Increases Protein Biosynthesis Yields

“…As a model, we explored this strategy in the context of cell-free protein synthesis (CFPS). CFPS is an attractive approach to produce proteins in vitro without the need to maintain cell growth. , In recent years, CFPS has matured to impact a variety of applications in diagnostics, biomanufacturing, and educational kits, among others. − Typically, the ribosome-containing lysates (S12 lysates) for CFPS are made from bacterial strains harboring multiple rRNA operons, producing a lysate with a heterogeneous ribosome pool (Figure A). Here, we sought to assess if protein synthesis could be increased by creating CFPS-capable lysates that do not contain ribosomes derived from these deleterious operons (e.g., operon D).…”

Validation of Cell-Free Protein Synthesis Aboard the International Space Station

“…Since both designs of biosensors are modular, they can be modified to detect other biochemically significant analytes, such as environmental markers, toxins, food and water contaminants, and viral, bacterial, and fungal nucleic acids. − To further streamline the use of these biosensors, the plasmids could be freeze-dried within the cell-free pellet, requiring just the addition of the environmental or biological sample in order to rehydrate the pellet and begin analysis. Given the extended shelf life, potential room temperature storage, and reduced weight and volume of lyophilized samples, lyophilization of the plasmid sample would be an ideal preparation mode of CFPS platforms designed for space. , Additionally, arrays or multiplexed systems of lyophilized CFPS biosensors producing analyte-specific wavelengths of fluorescence could be developed for the high-throughput detection of analytes.…”

Deconstructing synthetic biology across scales: a conceptual approach for training synthetic biologists

“…Fortunately, important first steps to developing new training approaches are beginning to happen with the emergence of new undergraduate opportunities and PhD programs in synthetic biology. For high school students and undergraduates, experiential learning opportunities have emerged to facilitate hands-on learning, such as BioBits Kits 28 – 31 , the ODIN marketplace for genetic engineering supplies 32 , BioBuilder 33 , and others 34 , 35 . In addition, opportunities such as the international Genetically Engineered Machines (iGEM) competition, the Build-a-Genome Course 36 , and the Cold Spring Harbor Summer Course in Synthetic Biology have paved the way to explore synthetic biology and this integration of disciplines.…”