Funded by a grant from the National Science Foundation: Future Manufacturing #2228971.
Synthetic biology and biomanufacturing are buzzwords of an emerging bioeconomy based on our increasing ability to manipulate living systems. This workshop will equip bioscience educators with the knowledge, skills, and resources to introduce students to a simple workflow to manufacture bacteriophages (phages) with new host specificity, illustrating how biomolecules are adapted to interact with cell-surface receptors.
Discover the cutting-edge cell-free transcription-translation (TXTL) system for in vitro phage biomanufacturing with instructors from the University of Minnesota and Cold Spring Harbor Laboratory. This workshop emerges from a distinguished lineage of bacteriophage research that began at CSHL and provided the first tools to explore the molecular mechanics of living cells. The “Phage Course,” founded at CSHL in 1945 by Max Delbrück and Salvador Luria, trained the first two generations of molecular biologists. Al Hershey and Martha Chase’s “blender experiment,” conducted at CSHL in 1952, provided conclusive evidence that DNA is the molecule of heredity. Delbrück, Luria, Hershey shared the 1969 Nobel Prize for this seminal work.
Bacteriophages are being rediscovered as powerful tools to meet the pharmaceutical challenge of untreatable, multi-drug resistant bacterial infections. In the food industry, bacteriophages prevent formation of biofilms on equipment surfaces, sanitize fresh fruits and vegetables, and extend the shelf life of packaged foods. Using phages to treat bacterial infections in livestock increases resilience and helps eliminate the reservoir of antibiotic-resistant bacteria. Phages also provide a virtually limitless source of bioactive materials that are increasingly exploited in biotechnology, nanotechnology, and bioremediation.
Phages are used in education as examples of simple genetic systems. The SEA-PHAGES Program of the Howard Hughes Medical Institute annotates phage genomes, and is one of the most widely implemented infrastructures for course-based undergraduate research experiences (CUREs). TXTL is a logical next step for students who have been exposed to phage and/or bacterial genetics, providing them an opportunity to explore the use of phages in biomanufacturing.
Workshop participants will conduct hands-on experiments to express reporter genes and whole phage genomes in vitro, using a cell-free extract. Participants will complete an entire workflow to engineer wild-type T7 phage to infect a new E. coli host. This begins with long PCRs to amplify several fragments of the T7 genome, plus PCR mutagenesis of the tail fiber gene. The PCR products are then assembled and packaged as complete T7 genomes using the cell-free TXTL system. Spotting assays compare the host range of wild-type and mutant T7 phages. Participants will then conduct nanopore DNA sequencing, delivering same-day results to investigate point mutations in the tail fiber gene that account for infectivity.
A $400 stipend will be provided. Travel funds are available.
Vincent Noireaux, Ph.D. is Professor of Synthetic Biology and Biological Physics at the University of Minnesota. His lab uses cell-free transcription-translation (TXTL) to produce proteins in vitro and to engineer biochemical systems such as gene circuits, phages, and synthetic cells. Dr. Noireaux earned a BS in Applied Physics from the University of Tours (France), his Ph.D. at the Curie Institute in Paris, and he did a postdoc at The Rockefeller University in New York City. He is co-instructor of the influential Cold Spring Harbor Laboratory postgraduate training course on Synthetic Biology.
Steven Bowden, Ph.D. is Assistant Professor of Food Safety Microbiology at the University of Minnesota. His lab studies foodborne pathogens such as Salmonella, Listeria monocytogenes, and Escherichia coli, and the use of bacteriophages to detect and control these bacteria. Dr. Bowden earned a BSc in Biochemistry from the University of Birmingham (UK) and his Ph.D. at the University of Cambridge (UK). He held postdoctoral positions at the Quadram Institute (UK), University of Cambridge (UK), Nara Institute of Science and Technology (Japan), and University of Minnesota.
Anna Feitzinger, Ph.D. is Assistant Director for Science of the DNA Learning Center of Cold Spring Harbor Laboratory, a science center devoted entirely to public genetics education. She applies her expertise in quantitative biology and biochemistry to bridging the gap in bioinformatics education at the pre-college and college level. She is spearheading the use of Oxford Nanopore sequencing in education, with the dream that sequencing technology will be available to students in classrooms worldwide. She received a B.A. in Chemistry from the City University of New York, Hunter College, and a Ph.D. in Biology at University of California, Davis.
Dave Micklos, D.Sc. is founder and Executive Director of the DNA Learning Center of Cold Spring Harbor Laboratory, which encompasses five science outreach labs in metro New York/New Jersey totaling 43,000 ft2. Each year, more than 500,000 students use textbooks or commercial lab kits developed by the DNALC, analyze DNA sequences using DNALC bioinformatics tools, or do experiments at the DNALC or an allied science center. Dave is a Fellow of the American Association for the Advancement of Science, and his work has been recognized with prizes from the Genetics Society of America, Charles A. Dana Foundation, American Society for Cell Biology Award, and Science Magazine.
6201 Winnetka Avenue, Woodland Hills, CA 91371
Funded by a grant from the National Science Foundation: Future Manufacturing #2228971.
