Life Sciences Commons^™

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 …

Genetic Tools Towards A Synthetic Biology Approach For Whole Mitochondrial Genome Engineering, Ryan R. Cochrane

Electronic Thesis and Dissertation Repository

Synthetic biology is an interdisciplinary research field that standardizes and repurposes biological components to better understand life and solve complex problems. As synthetic biology has developed, the goal has become to generate fully controllable biological systems through whole genome engineering (WGE), the cumulation of standardized genome engineering, and DNA delivery methods. In eukaryotes, genetic tools for WGE are limited to the nucleus and present a need to expand to include mitochondria, which maintains its own unique genome. The work presented here begins developing the resources needed to enable whole mitochondrial genome engineering.

First, to standardize mitochondrial genome engineering protocols, I …