Fig. 6.

Application of the Lightning GTR-CRISPR for free fatty acids production. a Schematic representations of lipid metabolism in S. cerevisiae. Gene names in red represent deletion targets by the Lightning GTR-CRISPR system. G3P glyceraldehyde 3-phosphate, LPA lysophosphatidic acid, PA phosphatidic acid, DAG diacylglycerol, TAG triacylglycerol, SE sterol ester, PI phosphatidylinositol, PC phosphatidylcholine, PE phosphatidylethanolamine, PS phosphatidylserine. FAA1 and FAA4 encode fatty acyl-CoA synthetases; POX1 encodes a fatty acyl-CoA oxidase; PAH1, DPP1, and LPP1 encode phosphate phosphatases, which dephosphorylate PA to yield diacylglycerol; and ARE1 and ARE2 encode acyl-CoA:sterol acyltransferases. b Free fatty acid and total fatty acid production in yeast deletion strains. Free fatty acids (intracellular and extracellular) and total fatty acids (including fatty acid chains in lipids: TAGs, SEs, phospholipids, etc.) were quantified when the strains were grown for 72 h in minimal medium containing 2% glucose. The data points of bar charts represent mean averages of indicated fatty acids. The error bars indicate the standard deviations of three biological replicates. Source data are provided as a Source Data file