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Full-Text Articles in Life Sciences
Developing Regulated Crispr Systems To Control Bacterial Microbiomes, Gregory M. Pellegrino
Developing Regulated Crispr Systems To Control Bacterial Microbiomes, Gregory M. Pellegrino
Electronic Thesis and Dissertation Repository
Changes to the human microbiome’s composition and metabolome are associated with numerous diseases and alterations to xenobiotic metabolism. As such, targeting the human microbiome is an increasingly popular option for therapeutic interventions. However, traditional therapies that target the microbiome such as antibiotics lack specificity, which can affect the beneficial species of the microbiome and cause adverse health outcomes such as the rise of antimicrobial-resistant bacteria. Therefore, the research and development of specific, targeted antimicrobial therapies is crucial to effectively treating microbiome dysbioses.
CRISPR and CRISPRi provide easily modifiable, RNA-guided mechanisms mediated by the Cas9 or dCas9 enzymes to induce sequence-specific …
Characterization Of Dna Regulatory Elements Of The Highly Abundant Secreted Protein 1 (Hasp1) Promoter And Analysis Of Transcriptomic Responses Under Phosphate Depletion In The Diatom Phaeodactylum Tricornutum, Mahsa Farmanbar
Electronic Thesis and Dissertation Repository
Phosphorus (P) is essential for all life. The bioavailability of phosphorus in oceans impacts diatoms like Phaeodactylum tricornutum (P. tricornutum). P. tricornutum is extensively used in transcriptomic studies to understand pathways involved in P-acquisition. However, activation mechanisms and roles of regulatory elements in P-acquisition responses remains unclear. Here, I deleted predicted phosphate regulatory sites in P.tricornutum’s HASP1 promoter to create different HASP1-eGFP constructs. Under P-depletion, two constructs showed increased eGFP secretion. Additionally, a HASP1 knockout strain was grown under different phosphorus sources to determine if HASP1 is a phytase. Cells grown in full phosphorus utilized organic …
Advances In Phaeodactylum Tricornutum Nuclear Engineering, Mark Pampuch
Advances In Phaeodactylum Tricornutum Nuclear Engineering, Mark Pampuch
Electronic Thesis and Dissertation Repository
The marine diatom Phaeodactylum tricornutum has the potential to become an excellent platform for the sustainable production of valuable compounds and pharmaceuticals, but currently large-scale engineering of this organism remains a challenge due factors like inefficient genetic transformation protocols and a lack of accurate genomic data. This thesis addresses these two bottlenecks by (i) optimizing an electroporation protocol to P. tricornutum and (ii) remapping genomic data from a scaffolded genome assembly to a telomere-to-telomere genome assembly. An optimized transformation protocol was developed that could consistently transform blunt-ended and DNA with overhangs and yielded up to 1000+ colony forming units per …
Functionalizing Conjugative Systems To Deliver Crispr Nucleases For Targeted Bacterial Killing, Thomas A. Hamilton
Functionalizing Conjugative Systems To Deliver Crispr Nucleases For Targeted Bacterial Killing, Thomas A. Hamilton
Electronic Thesis and Dissertation Repository
The interactions between humans and microbes are intimately important to human health, with both commensal and pathogenic bacteria affecting homeostasis and disease. Increasing concern over antibiotic resistance in bacterial pathogens represents a significant threat to human health, and use of traditional antibiotics to treat infections can be detrimental to commensal bacteria as well as pathogens, demonstrating a need for more specific antibacterial reagents. RNA-guided CRISPR nucleases, which can target and cleave genomes of interest, are a potential tool for specific bacterial targeting. A key limitation to the use of CRISPR antimicrobials is effective and robust delivery to the target bacteria. …