Rapid optimization of protein function in mammalian cells via microbe-independent deep assembly and screening
Wu, Y.; Wang, P.; Liu, L. X.; Gao, C.; Qin, Q.; Hageman, M.; Kirkland, T. A.; Su, Y.; Lin, M. Z.
Show abstract
Random mutagenesis and deep mutational scanning (DMS) are widely used to optimize proteins by oversampling large libraries in microbial cells, selecting cells expressing favorable variants, and sequencing to identify enriched variants. However, these methods are slow and costly, require effort to establish selection methods for a given protein function, and do not yield data on lower-performing variants. Here, we describe Microbe-Independent Deep Assembly and Screening (MIDAS), a rapid, high-throughput method for optimizing protein function directly in mammalian cells. As a demonstration, we applied MIDAS to improve a newly designed neurotransmitter bioluminescent indicator (NeuBI) for acetylcholine (ACh). MIDAS systematically optimized interdomain linkers, identified mutational hotspots, and exhaustively scanned amino acid combinations, in each case relating specific sequences to protein performance. MIDAS-optimized variants exhibited improved performance in vivo, highlighting the potential of MIDAS for improving protein function in mammalian systems.
Matching journals
The top 5 journals account for 50% of the predicted probability mass.
Similar papers in this journal
Similar papers in this journal
Similar papers in this journal
- Fluorescence lifetime enables high-resolution analysis of neuromodulator dynamics across time and animals 96%
- Bright and sensitive red voltage indicators for imaging action potentials in brain slices and pancreatic islets 95%
- Multiplexed whole animal imaging with reversibly switchable optoacoustic proteins 95%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.