Through funding from a National Science Foundation Advanced Technological Education (NSF-ATE) grant, we have developed an innovative laboratory exercise and curriculum on CRISPR gene editing. The kit exercise provides the ability to perform state of the art CRISPR gene editing laboratory procedures, giving students at the high school and introductory college course level the ability to learn the science behind current gene editing efforts without the need for elaborate materials or equipment. At its base, this exercise enables students to practice CRISPR-directed gene editing using a mammalian cell-free extract, a plasmid bearing the lacZ gene and an appropriate CRISPR-Cas (Cas9 or Cas12a) complex which together catalyze a genetic deletion or insertion/replacement of a new sequence when a single-stranded DNA donor template is provided. Both genetic knockout and genetic knock-in reactions are active in the instructional kit. Importantly, this procedure does not lead to the direct genetic modification of bacterial cells, which could lead to GMO issues of safety concerns; instead, bacterial cells are simply used as the readout after a standard transformation.
The laboratory exercise models and mimics gene editing in mammalian cells, which is the major interest of over 90% of the gene editing activities in the field, without the need for special equipment including tissue culture hoods, CO2 incubators and any sort of microscopy. The reaction can be broken down into individual teachable modules and is direct, rapid, cost-effective and takes advantage of standard molecular biology reagents and equipment that are likely present in many high school biotechnology program labs and most community college instructional laboratories. Curriculum enhances standard biology syllabi content of chemical information flow, genetic transformation, lacZ function, DNA function, etc.
SPEAKERS:Eric Kmiec, Ph.D. Director, Gene Editing Institute, Helen F. Graham Cancer Center & Research Institute, Christiana Care and John McDowell, Ph.D. Instructor Biology/Chemistry Department, Delaware Technical Community College