Integration of Authentic Research into an Undergraduate Laboratory Course: Design and Synthesis of a Gene Therapy Vector

Abstract

Undergraduate Biology students often graduate without exposure to authentic research experiences. Laboratory courses often follow a one or two week fail‐proof experiment resembling a cookbook recipe, lacking the uncertainty of genuine research. Techniques in genetics and molecular biology cover a wide array of skills that are essential to succeed in a biotechnological laboratory today. Can we design a lab course based on the teaching of concepts while imparting the skills and applications of modern techniques to undergraduate students? In our experimental laboratory course, we focus on the implementation of genetics and molecular biology lab techniques, thus providing students with theoretical concepts and laboratory skills. We prepare students to carry‐out scientific protocols that can be applied to a future workforce setting. Students are immersed in an 11‐week series of labs with the objective to use molecular cloning to make a gene therapy vector; therapies are designed to inhibit the overexpression of oncogenes in brain tumors. Students in the laboratory are introduced to PubMed and Genbank to research the background of the target gene. Students use DNA analysis software, Serial Cloner, as an interactive tool to evaluate DNA sequences and visualize therapy design, with each student generating their own therapy vector. Students isolate and purify DNA using a plasmid purification protocol, and measure quantity and quality using a spectrophotometer. They are introduced to molecular cloning and the value of master mixes, beginning with the digestion of their DNA plasmid using restriction enzymes. Students, making their own buffers and gels, perform gel electrophoresis and subsequently cut‐out the appropriate bands. Students purify bands and clean‐up DNA with magnetic beads, preparing samples for the next step. Students perform DNA ligation with proper controls and transform ligations into DH5‐α super competent cells. To verify successful transformation, students make Luria Broth ampicillin plates and spread transformed bacteria onto plates; ligation efficiency is measured based on bacterial colony count. The students then use colony PCR and agarose gel analysis to verify successful ligations. At the end of the semester, students compile results from each step during the 11‐week lab series and integrate learning from the lecture to synthesize a comprehensive lab report. Student evaluation is based on connecting lecture concepts to laboratory experiments, including clearly stating lab objectives, writing protocols in own words, accurately recording results in notebook (in pen), labeling of figures, analysis of data, and comprehension of results. Properly recording and understanding results is more highly valued over expected results. Successful experiments carried‐out in the course are used to contribute to authentic research projects. The goal of this lab course is to integrate authentic research into the undergraduate learning experience, preparing students for graduate school and employment in a modern biotechnological lab; establishing the next generation of scientist.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.