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dc.contributor.authorDiCarlo, James Edwarden_US
dc.date.accessioned2016-01-05T18:22:41Z
dc.date.available2016-01-05T18:22:41Z
dc.date.issued2015
dc.identifier.urihttps://hdl.handle.net/2144/13712
dc.description.abstractThe field of synthetic biology seeks to use design principles of life to create new genes, organisms and populations to both better understand biology as well as generate species with useful properties. Budding yeast has been a workhorse for synthetic biology, as well as an important model organism in the broader fields of molecular biology and genetics. This thesis aimed to create genome engineering tools for the manipulation of genomes, with direct applications in yeast. I focused developing high-throughput and highly efficient methods for making genomic modifications in yeast to allow for the generation of large libraries of precisely modified yeast genomes. By manipulation of endogenous DNA recombinases and mismatch repair enzymes in yeast, we were able to develop an oligonucleotide only method for genome engineering to generate libraries as large as 10^5 individuals with a frequency of modification as high as 1%. Additionally, we validated the use of RNA-guided CRISPR/Cas9 endonucleases to make changes in yeast genomes, resulting in frequencies of genome modification >90% in transformed populations. We further optimized this method to generate larger libraries as high as 10^5 individuals and explored a proof of concept epistasis experiment involving thermotolerance. Lastly, the propagation of changes to successive generations is useful when engineering organisms on the population level. To this end we explored the use of RNA-guided gene drives to bias inheritance in S. cerevisiae. We show that inheritance of these selfish elements can be biased to over 99% and is reversible.en_US
dc.language.isoen_US
dc.subjectBiomedical engineeringen_US
dc.subjectCas9en_US
dc.subjectCRISPRen_US
dc.subjectMAGEen_US
dc.subjectYeasten_US
dc.subjectGene driveen_US
dc.subjectGenome engineeringen_US
dc.titleEngineering yeast genomes and populationsen_US
dc.typeThesis/Dissertationen_US
dc.date.updated2015-10-28T01:09:01Z
etd.degree.nameDoctor of Philosophyen_US
etd.degree.leveldoctoralen_US
etd.degree.disciplineBiomedical Engineeringen_US
etd.degree.grantorBoston Universityen_US


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