Progress in the biological sciences relies on the availability of appropriate tools and strategies; in the areas of understanding and constructing multicellular systems, better methods of gene expression control are needed. Here, we work towards two approaches to controlling gene expression for applications related to multicellular coordination and pattern formation. First, we characterize the use of 2A self-cleaving peptides for polycistronic expression in Dictyostelium discoideum, a powerful model system for investigating the onset of coordinated behavior in a cell population. We also make novel observations about various factors that affect gene expression levels, which would inform future decisions on the use of 2A peptides in this system. Second, we examine the use of antisense RNA in mammalian cells for gene repression from an input that simultaneously activates other genes. We show that in transient overexpression systems, antisense RNA produced in trans can be a more effective repressor than convergent promoters and other well-established RNA-based methods for gene repression, and we explore ways in which this system could be improved for stable gene circuits. Overall, our results advance our understanding of available tools for gene expression control and the conditions under which they are appropriate to use.
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/49246 |
| Date | 11 September 2024 |
| Creators | Zhu, Xinwen |
| Contributors | Wong, Wilson W |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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