[P007] Poster

12:10 PM - 1:00 PM JST(3:10 AM - 4:00 AM UTC)

[P04-449] Dual-channel optogenetics in yeast for multiplexed light-based control of cellular processes and pathways

○LINUS TAN TAN^1,2, Zhangyuan Lin^1,2, Jing Yui Yeoh^1,2, Jingyun Zhang^1,2, Chueh Loo Poh^1,2 (1. National University of Singapore, Department of Biomedical Engineering (Singapore), 2. NUS Synthetic Biology for Clinical and Technological Innovation (Singapore))

Keywords:

Optogenetics,S. cerevisiae,Biomanufacturing,Genetic circuit

Optogenetics allows the expression of genes to be controlled dynamically, with spatial-temporal resolution and in a manner that is orthogonal to native cell metabolism and has found great utility in metabolic engineering and biomanufacturing. S. cerevisiae is an industrially relevant model organism that is used in many applications, but due to the large number of genes required and issues relating to cross-activation between different colours, optogenetics for different wavelengths of light have not been multiplexed in S. cerevisiae. In this paper, we develop a compact red light responsive optogenetic system for S. cerevisiae that requires only a single gene and no exogenous cofactors. Through engineering modular protein domains, we reduce the cross-activation of our system by blue light. We integrate our novel red light optogenetic system with EL222 blue light optogenetics to establish the first dual channel optogenetics in S. cerevisiae and demonstrate its utility for engineering biology through the light-based control of flavonoid luteolin synthesis and flocculation for ease of product extraction. We also demonstrate our system’s potential for the biomanufacturing of living materials by producing the first dual-coloured optogenetic patterns using S. cerevisiae. This work expands optogenetic applications in S. cerevisiae from single-light to multi-light systems, introducing the potential to multiplex different colours of light for dynamic, orthogonal control of separate cell processes.