{"671863":{"#nid":"671863","#data":{"type":"event","title":"2024 Suddath Award Winner Presentation","body":[{"value":"\u003Cp\u003E\u003Cstrong\u003E\u003Ca href=\u0022https:\/\/sites.gatech.edu\/blazecklab\/lab-members\/\u0022 target=\u0022_blank\u0022\u003EAndrew Cazier\u003C\/a\u003E, Ph.D. Candidate\u003C\/strong\u003E\u003Cbr \/\u003E\r\n2024 Suddath Award Winner\u003Cbr \/\u003E\r\nJohn Blazeck, Ph.D., Advisor\u003Cstrong\u003E\u0026nbsp;\u003C\/strong\u003E\u003C\/p\u003E\r\n\r\n\u003Cp\u003E\u003Cstrong\u003EABSTRACT\u003C\/strong\u003E\u003Cbr \/\u003E\r\nThe yeast Saccharomyces cerevisiae is an excellent host for directed evolution experiments, but classical methods to introduce mutations can be time-consuming or inefficient. To address these limitations, we have built a CRISPR-based, yeast diversifying base editor (yDBE) that can generate DNA mutant libraries in situ. This system can cause substantial and diverse mutations in a 100-bp window near each targeted locus and can target multiple loci simultaneously. To demonstrate the capabilities of this platform, we have successfully applied yDBE to rapidly enhance and isolate antibody sequences with up to 100-fold higher affinity and greatly increase the strength of a weak, exogenous promoter. We will highlight our optimization strategies which significantly improved the mutation rate or breadth, including 1) applying an improved deaminase variant, 2) modifying the CRISPR guide RNA (gRNA) scaffold, and 3) building multi-gRNA cassettes. We will also outline ongoing experiments to employ yDBE such as to isolate high affinity antibodies from a na\u00efve library. Ultimately, we will show that our system is a rapid, inexpensive, and easily-incorporated tool for directed evolution in yeast.\u003C\/p\u003E\r\n\r\n\u003Cp\u003E\u003Cbr \/\u003E\r\n\u003Cem\u003EThe F. L. \u0022Bud\u0022 Suddath and Frances \u0022Lee\u0022 Gafford Suddath Fellowship Award was established by family and friends in recognition of Bud Suddath\u0027s contributions as a Georgia Tech faculty member and to support Bud and Lee\u0027s shared interest in graduate education in the fields of biology, biochemistry and biomedical engineering. The fellowship award is given annually to Georgia Tech Ph.D. students, typically within one year of graduation, who have demonstrated significant achievement in research of fundamental importance in the fields of biology, biochemistry, or biomedical engineering and are working in laboratories affiliated with Georgia Tech\u0027s Institute of Bioengineering and Biosciences (IBB).\u003C\/em\u003E\u003C\/p\u003E\r\n","summary":"","format":"limited_html"}],"field_subtitle":"","field_summary":[{"value":"\u003Cp\u003E\u201cTargeted DNA Diversification and Directed Evolution in Saccharomyces Cerevisiae Using a CRISPR Base Editor\u201d\u003C\/p\u003E\r\n","format":"limited_html"}],"field_summary_sentence":[{"value":"\u201cTargeted DNA Diversification and Directed Evolution in Saccharomyces Cerevisiae Using a CRISPR Base Editor\u201d - Andrew Cazier, Ph.D. Candidate"}],"uid":"35486","created_gmt":"2024-01-04 13:39:30","changed_gmt":"2024-01-04 13:40:45","author":"Christina Wessels","boilerplate_text":"","field_publication":"","field_article_url":"","field_event_time":{"event_time_start":"2024-03-14T11:00:00-04:00","event_time_end":"2024-03-14T12:00:00-04:00","event_time_end_last":"2024-03-14T12:00:00-04:00","gmt_time_start":"2024-03-14 15:00:00","gmt_time_end":"2024-03-14 16:00:00","gmt_time_end_last":"2024-03-14 16:00:00","rrule":null,"timezone":"America\/New_York"},"location":"Petit Biotech Building (IBB), Suddath Seminar Room 1128, 315 Ferst Drive","extras":[],"groups":[{"id":"1292","name":"Parker H. Petit Institute for Bioengineering and Bioscience (IBB)"}],"categories":[],"keywords":[{"id":"187423","name":"go-bio"}],"core_research_areas":[],"news_room_topics":[],"event_categories":[{"id":"1795","name":"Seminar\/Lecture\/Colloquium"}],"invited_audience":[{"id":"78761","name":"Faculty\/Staff"},{"id":"177814","name":"Postdoc"},{"id":"78771","name":"Public"},{"id":"174045","name":"Graduate students"},{"id":"78751","name":"Undergraduate students"}],"affiliations":[],"classification":[],"areas_of_expertise":[],"news_and_recent_appearances":[],"phone":[],"contact":[{"value":"\u003Cp\u003E\u003Ca href=\u0022mailto:connect@ibb.gatech.edu\u0022\u003Econnect@ibb.gatech.edu\u003C\/a\u003E - event inquiries\u003C\/p\u003E\r\n","format":"limited_html"}],"email":[],"slides":[],"orientation":[],"userdata":""}}}