Shotgun genetic engineering enables high-throughput metabolic design in mammalian cells
Researchers developed shotgun genetic engineering (SGE) to overcome the combinatorial complexity of metabolic engineering in mammalian cells. By delivering barcoded, small DNA constructs into millions of cells simultaneously, SGE allows each cell to act as an independent experiment. This approach successfully engineered essential amino acid biosynthesis in Chinese hamster ovary (CHO) and Jurkat cells, achieving near-wild-type growth rates without valine or isoleucine. The method identified optimal mitochondrial localization and gene stoichiometry, outperforming previous rational design strategies and generating data for machine learning models.