Engineering gene overlaps to sustain genetic constructs in vivo

• DOI: 10.1371/journal.pcbi.1009475
• Publication Date: 2021-10-09T23:56:06Z
• Authors: Antoine L. Decrulle; Antoine Frénoy; Thomas A. Meiller...; Aude Bernheim; Chantal Lotton; Arnaud Gutierrez; Ariel B. Lindner
• Keywords: Synthetic Biology; Negative selection; Genetic Fitness; Gene selection

Evolution is often an obstacle to the engineering of stable biological systems due selection mutations inactivating costly gene circuits. Gene overlaps induce important constraints on sequences and their evolution. We show that these can be harnessed increase stability genes by purging loss-of-function mutations. combine computational synthetic biology approaches rationally design overlapping reading frame expressing essential within existing protect. Our algorithm succeeded in creating frames 80% E. coli genes. Experimentally, scoring both such construct, we found a significant fraction impacting protect have deleterious effect gene. Such overlap thus protects from removal natural associating benefit this with larger or even lethal cost. In our constructs, converts many possible mutants into evolutionary dead-ends, reducing potential system increasing its over time.