Large scale microfluidic CRISPR screening for increased amylase secretion in yeast
S. Johansson, Thierry Dulermo, Cosimo Jann, Justin Smith, Anna Pryszlak, Georges Pignède, Daniel Schraivogel, Didier Colavizza, Thomas Desfougères, Christophe Rave, Alexander Farwick, Christoph A. Merten, Kevin Roy, Wei Wu, Lars M. Steinmetz
Abstract
, we identified 345 genes for which an increase or decrease in gene expression resulted in increased secretion of α-amylase. Our results show that modulating the expression of genes involved in the trafficking of vesicles, endosome to Golgi transport, the phagophore assembly site, the cell cycle and energy supply improve α-amylase secretion. Besides protein-coding genes, we also find multiple long non-coding RNAs enriched in the vicinity of genes associated with endosomal, Golgi and vacuolar processes. We validated our results by overexpressing or deleting selected genes, which resulted in significant improvements in α-amylase secretion. The advantages, in terms of precision and speed, inherent to CRISPR based perturbations, enables iterative testing of new strains for increased protein secretion.