| Products/Services Used | Details | Operation |
|---|---|---|
| Codon Optimization | Generation and Screening of a Combinatorial Library to Identify Improved NfsA Variants Based on the above results, a combinatorial gene library was generated (GenScript) to encode all 512 possible combinations of the 10 residue substitutions at the 8 identified positions and the corresponding wild-type residues (Figure S1).... Combinatorial Library Construction Eight identified positions (I5T, S41Y, E99G, L103M, K222E, R225x [x = A, G, P]), F227S, and L229V were chosen for inclusion in a synthetically constructed mutant gene library (GenScript; Figure S1), which, through the use of redun- dancy codons, was able to code for all possible 512 combinations of these mu- tations. | Get A Quote |
Gene-directed enzyme-prodrug therapy (GDEPT) is a promising anti-cancer strategy. However, inadequate prodrugs, inefficient prodrug activation, and a lack of non-invasive imaging capabilities have hindered clinical progression. To address these issues, we used a high-throughput Escherichia coli platform to evolve the multifunctional nitroreductase E. coli NfsA for improved activation of a promising next-generation prodrug, PR-104A, as well as clinically relevant nitro-masked positron emission tomography-imaging probes EF5 and HX4, thereby addressing a critical and unmet need for non-invasive bioimaging in nitroreductase GDEPT. The evolved variant performed better in E. coli than in human cells, suggesting op... More
