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Efficient and Rapid Discovery of Novel Temperature Sensitive Alleles with the Onyx ® Digital Genome Engineering Platform

Temperature sensitive (TS) alleles are mutations in essential genes that allow for growth at permissive temperatures and restrict or reduce growth at non-permissive temperatures. These conditional mutations are useful in academic settings for basic study of essential genes, and they are useful in bio-industrial settings as many essential genes are central to the production of high-value compounds. While critical for the study and use of essential genes, TS alleles are notoriously difficult to find. During this webinar, we will showcase how we leverage the precise, trackable genome editing of the Onyx Platform to discover novel TS alleles in yeast and E. coli. Following genome engineering, selection experiments have led to the identification of many potential TS alleles. Initial validation experiments are proving that many of these potential hits are novel TS alleles that show distinct growth properties when comparing permissive and non-permissive growth temperatures. These novel conditional mutants will function as invaluable tools for future studies and industrial applications of essential genes. This work highlights the effectiveness of Inscripta’s Digital Genome Engineering platform for novel discoveries in both yeast and E. coli.