4 research outputs found
Tuning DNA binding and gene expression using zinc finger proteins and engineered promoters
Synthetic biology provides an ideal approach to build functional biological devices by assembling biological parts. Using synthetic biology, efficient control of gene regulation may be achieved to a degree that is not possible using natural genetic structures.1 However, previous studies on promoter engineering have focused on natural transcription factors (TFs),2 including the lac repressor, which produces a switch-like “all-or-none” response.3 In this project, we worked to develop a new system for transcriptional control based on tunable synthetic TFs, which are designed to yield programmable linear responses in gene expression. To accomplish this, we used zinc finger proteins (ZFPs) as regulators of engineered promoters assayed by green fluorescent protein (GFP) as a fluorescent transcriptional reporter probe. In particular, we designed strong-binding three finger ZFPs as proof-of-principle regulatory elements, with the intention of moving to weaker binding two finger ZFPs and the addition of the accessory binding module PAR (part of the protein Adr1).4 To generate engineered promoters, we integrated ZFP binding sites into known promoters of varying strength. To analyze the engineered activity of each promoter, we cultured E. coli cells transformed with plasmids containing sequences for both ZFP production and our engineered promoters and measured the resulting fluorescence intensity. In this way, we constructed a novel method for tuning gene expression as well as testing the DNA binding affinity of synthetic TFs. We anticipate that this general approach could be used in the future for designing and characterizing synthetic TFs for gene therapy and gene regulation applications
Improving motivation and engagement in core engineering courses with student teams
Team-based projects are common in capstone engineering design courses and increasingly common in first-year engineering programs. Despite high enrollments and budget cutbacks affecting many programs, second- and third-year students can also benefit from team-based project experiences, which motivate them to succeed in engineering and prepare them for a globally competitive workforce. My dissertation research demonstrates that team design projects can be incorporated into the curricula of engineering departments, and these projects result in positive affective outcomes for students. Using ABET outcomes and Self Determination Theory (SDT) as the background for my studies, I investigated students' confidence, motivation, and sense of community after experiencing team design projects in two different engineering departments at a large public institution. In the first study, I used a sequential mixed methods approach with a primary quantitative phase followed by an explanatory qualitative phase to evaluate a chemical engineering program that integrated team design projects throughout the curriculum. The evaluation methods included a survey based on desired ABET outcomes for students and focus groups to expand on the quantitative results. Students reported increased confidence in their design, teamwork, and communication skills after completing the projects. In my second and third studies, I used qualitative interviews based on SDT to explore student motivation in an electrical and computer engineering course redesigned to support students' intrinsic motivation to learn. SDT states that intrinsic motivation to learn is supported by increasing students' sense of autonomy, competence, and relatedness in regard to their learning. Using both narrative inquiry and phenomenological methodologies, I analyzed data from interviews of students for mentions of autonomy, competence, and relatedness as well as course events that were critical in changing students' motivation. Analysis revealed that individual choice, constructive failures, and a strong sense of community in the classroom were critical to moving students toward intrinsic motivation. Further, community building through team experiences characterized the essence of the student experience in the course. My research highlights a need for better quantitative measures of students' affective outcomes, specifically motivation, in the context of a single course. Based on the results of my studies, SDT should be reevaluated in terms of possible interdependencies between autonomy, competence, and relatedness, and how the social context of large engineering courses may create a deeper need for supporting relatedness
Improving Motivation and Engagement in Core Engineering Courses with Student Teams
Team-based projects are common in capstone engineering design courses and increasingly common in first-year engineering programs. Despite high enrollments and budget cutbacks affecting many programs, second- and third-year students can also benefit from team-based project experiences, which motivate them to succeed in engineering and prepare them for a globally competitive workforce. My dissertation research demonstrates that team design projects can be incorporated into the curricula of engineering departments, and these projects result in positive affective outcomes for students. Using ABET outcomes and Self Determination Theory (SDT) as the background for my studies, I investigated students’ confidence, motivation, and sense of community after experiencing team design projects in two different engineering departments at a large public institution. In the first study, I used a sequential mixed methods approach with a primary quantitative phase followed by an explanatory qualitative phase to evaluate a chemical engineering program that integrated team design projects throughout the curriculum. The evaluation methods included a survey based on desired ABET outcomes for students and focus groups to expand on the quantitative results. Students reported increased confidence in their design, teamwork, and communication skills after completing the projects. In my second and third studies, I used qualitative interviews based on SDT to explore student motivation in an electrical and computer engineering course redesigned to support students’ intrinsic motivation to learn. SDT states that intrinsic motivation to learn is supported by increasing students’ sense of autonomy, competence, and relatedness in regard to their learning. Using both narrative inquiry and phenomenological methodologies, I analyzed data from interviews of students for mentions of autonomy, competence, and relatedness as well as course events that were critical in changing students’ motivation. Analysis revealed that individual choice, constructive failures, and a strong sense of community in the classroom were critical to moving students toward intrinsic motivation. Further, community building through team experiences characterized the essence of the student experience in the course. My research highlights a need for better quantitative measures of students’ affective outcomes, specifically motivation, in the context of a single course. Based on the results of my studies, SDT should be reevaluated in terms of possible interdependencies between autonomy, competence, and relatedness, and how the social context of large engineering courses may create a deeper need for supporting relatedness.National Science Foundation / DUE-1140554U of I Onlythesi