Chappell, James ELoveless, Theresa B2024-05-212024-05-212024-052024-01-25May 2024Elizondo, Andrea Garza. Targeted Transcriptional Activation Using a CRISPR-Associated Transposon System. (2024). PhD diss., Rice University. https://hdl.handle.net/1911/116076https://hdl.handle.net/1911/116076Synthetic perturbation of gene expression is central to our ability to reliably uncover genotype-phenotype relationships in microbes. Here, I present a novel gene transcription activation strategy that uses the Vibrio cholerae CRISPR-Associated Transposon (CAST) system to selectively insert promoter elements upstream of genes of interest. Through this strategy, I show robust activation of both recombinant and endogenous genes across the Escherichia coli chromosome. I then demonstrate precise tuning of expression levels by exchanging the promoter elements being inserted. Finally, I demonstrate that CAST activation can be used to synthetically induce ampicillin-resistant phenotypes in E. coli.application/pdfengCopyright is held by the author, unless otherwise indicated. Permission to reuse, publish, or reproduce the work beyond the bounds of fair use or other exemptions to copyright law must be obtained from the copyright holder.CRISPR-Associated TransposonCRISPRTranscriptional ActivationThesis2024-05-21