A laboratory module that explores RNA interference and codon optimization through fluorescence microscopy using Caenorhabditis elegans

Abstract

AbstractAuthentic research experiences are beneficial to students allowing them to gain laboratory and problem-solving skills as well as foundational research skills in a team-based setting. We designed a laboratory module to provide an authentic research experience to stimulate curiosity, introduce students to experimental techniques, and promote higher-order thinking. In this laboratory module, students learn about RNA interference (RNAi) and codon optimization using the research organism Caenorhabditis elegans (C. elegans). Students are given the opportunity to perform a commonly used method of gene downregulation in C. elegans where they visualize gene depletion using fluorescence microscopy and quantify the efficacy of depletion using quantitative image analysis. The module presented here educates students on how to report their results and findings by generating publication quality figures and figure legends. The activities outlined exemplify ways by which students can acquire the critical thinking, data interpretation, and technical skills, which are beneficial for future laboratory classes, independent inquiry-based research projects and careers in the life sciences and beyond.SCIENTIFIC TEACHING CONTENTLearning GoalsGain experience working with C. elegansUnderstand the process of RNA interference and importance of codon optimizationLearn basic microscopy techniques and image analysisLearn how to properly use the scientific methodEnhance critical thinking skillsLearning ObjectivesStudents will be able to:Lab 1 and 2: Identify specific larval stages of C. elegansSynchronize C. elegans larvae using alkaline hypochlorite treatmentUnderstand codon usageFormulate hypotheses and design a controlled experimentLab 3 and 4: Acquire images using an epifluorescence microscopeEffectively communicate results and formulate conclusions from dataDescribe what RNAi is and how it affects gene expression/activityCalculate mean fluorescent intensity from acquired fluorescence micrographsPerform statistical tests to determine the significance of resultsGenerate publication quality figures and figure legends