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Browsing by Author "Sun, Wei"

Now showing 1 - 2 of 2
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    Developing a Novel Cell Surface RNA Detecting and Profiling Method via RNA Metabolic Labeling
    Brooks, Maxwell David (Virginia Tech, 2024-06-03)
    Cell surface RNA (csRNA) is a recent discovery in the field of RNA biology and has been implicated in playing important roles in many biological processes due to its extracellular properties. To understand the biogenesis, regulation, and function of csRNA, it is critical to develop methods to detect, isolate, and confidently characterize membrane-bound csRNA. Previously, csRNA has been profiled using methods based on cell membrane isolation that are expensive, laborious, and with unsatisfactory specificity and sensitivity . In this study, we use metabolic labeling and chemical cross-linking techniques to specifically label csRNA with biotin handles. We intended to use this technique for separating biotin-labeled csRNA from total RNA samples for characterization purposes. The primary materials that were used to label such csRNAs are 4-Thiouridine (4sU), an unnatural nucleotide analogue, and S-(2-aminoethyl)-ester-methanesulfonothioic-acid-biotin (MTSEA-biotin), a crosslinker designed specifically to label 4sU. By deploying these tools to cell lines such as HEK293T and HeLa, csRNA is detectable by Enhanced Chemiluminescent detection via Dot Blot. Furthermore, to separate biotin-labeled csRNA from total RNA, streptavidin-coated magnetic bead separation procedures could be used as a promising method for purifying csRNA from total RNA, for RNAseq characterization. This study highlights the processes of establishing the csRNA detection protocol and describes the current status and issues with developing the streptavidin-coated magnetic beads separation method.
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    The relationship among middle school students' motivation perceptions of science class, science identification and career goals
    Sun, Wei (Virginia Tech, 2018-06-04)
    This dissertation examined the extent to which pre-high school students' motivation-related perceptions of their science class affected their science identification, which sequentially affected their future science-related career goals. The MUSIC® Model of Motivation (Jones, 2009, 2018) includes five components (i.e., eMpowerment, Usefulness, Success, Interest, and Caring) and is designed to help teachers design instruction to promote students' motivation. Domain identification (Osborne and Jones, 2011) is a concept closely related to students' motivation and academic outcomes. In this study, data was collected from 311 pre-high school students and Structural Equation Modeling (SEM) analysis was conducted to test the structure pattern among the MUSIC model components, science identification, and science-related career goals. Results indicate that with three of the MUSIC model components (i.e., usefulness, success, and interest) significantly related to students' science identification, students' science identification was highly correlated to their science career goals. Moreover, this study demonstrated the structure patterns among the MUSIC model components and science identification varied by gender by conducting multi-group SEM analyses for a separate female sample (N = 161) and male sample (N = 150). Consistently, students' science identification was a strong predictor of their science career goals in both female and male groups. These findings are important for STEM educators because they indicate that it may be possible for teachers to impact students' science identification and career goals by focusing on students' perceptions of the MUSIC model components in science class. Moreover, these results contribute to the study of the large gender gap in STEM careers. Teachers can focus on specific teaching strategies and help female students develop their science identification in ways that lead to their long-term science-related career goals.