Transient overexpression of DNA adenine methylase enables efficient and mobile genome engineering with reduced off-target effects.

ABSTRACT: Homologous recombination of single-stranded oligonucleotides is a highly efficient process for introducing precise mutations into the genome of E. coli and other organisms when mismatch repair (MMR) is disabled. This can result in the rapid accumulation of off-target mutations that can mask desired phenotypes, especially when selections need to be employed following the generation of combinatorial libraries. While the use of inducible mutator phenotypes or other MMR evasion tactics have proven useful, reported methods either require non-mobile genetic modifications or costly oligonucleotides that also result in reduced efficiencies of replacement. Therefore a new system was developed, Transient Mutator Multiplex Automated Genome Engineering (TM-MAGE), that solves problems encountered in ot

SUBMITTER: Lennen RM 

PROVIDER: S-EPMC4770203 | biostudies-literature | 2016 Feb

REPOSITORIES: biostudies-literature