CRISPR-Cas9-assisted genome editing inE. colielevates the frequency of unintended mutations
Karl A. Widney,
Dong-Dong Yang,
Leo M. Rusch
et al.
Abstract:Cas-assisted lambda Red recombineering techniques have rapidly become a mainstay of bacterial genome editing. Such techniques have been used to construct both individual mutants and massive libraries to assess the effects of genomic changes. We have found that a commonly used Cas9-assisted editing method results in unintended mutations elsewhere in the genome in 26% of edited clones. The unintended mutations are frequently found over 200 kb from the intended edit site and even over 10 kb from potential off-tar… Show more
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