Simultaneous non-contiguous deletions using large synthetic DNA and site-specific recombinases.

ABSTRACT: Toward achieving rapid and large scale genome modification directly in a target organism, we have developed a new genome engineering strategy that uses a combination of bioinformatics aided design, large synthetic DNA and site-specific recombinases. Using Cre recombinase we swapped a target 126-kb segment of the Escherichia coli genome with a 72-kb synthetic DNA cassette, thereby effectively eliminating over 54 kb of genomic DNA from three non-contiguous regions in a single recombination event. We observed complete replacement of the native sequence with the modified synthetic sequence through the action of the Cre recombinase and no competition from homologous recombination. Because of the versatility and high-efficiency of the Cre-lox system, this method can be used in any organism where

SUBMITTER: Krishnakumar R 

PROVIDER: S-EPMC4132700 | biostudies-literature | 2014 Aug

REPOSITORIES: biostudies-literature