SUMMARY
Gene disruption has been drastically facilitated by recent genome editing tools. While the efficiency of gene disruption in cell culture has improved, clone isolation remains routinely performed to obtain fully mutated cells, potentially leading to artifacts due to clonal variances in cellular phenotypes. Here we report GENF, a highly efficient strategy to disrupt genes without isolating clones, which can be multiplexed. We obtained reliable lipidomics datasets from mutant cells generated with GENF, which was impossible when using clones. Through this, we found that an enzyme involved in congenital generalized lipodystrophy regulates glycerophospholipids with specific acyl-chains. We also demonstrate the possibility to dissect complex lipid co-regulatory networks and provide some common mechanisms explaining the adaptations to altered lipid metabolism. GENF is likely to contribute to all experimental setups affected by clonal variability and should be especially useful for -omics approaches.
Competing Interest Statement
The Department of Lipid Signaling, National Center for Global Health and Medicine, is financially supported by Ono Pharmaceutical Co., Ltd., Japan. The Department of Lipidomics, Graduate School of Medicine, The University of Tokyo, is funded by Shimadzu Corp., Japan.