Simultaneous non-contiguous deletions using large synthetic DNA and site-specific recombinases
作者： Radha Krishnakumar , Carissa Grose , Daniel H. Haft , Jayshree Zaveri , Nina Alperovich
DOI: 10.1093/NAR/GKU509
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摘要: Toward achieving rapid and large scale genome modification directly in a target organism, we have developed new engineering strategy that uses combination of bioinformatics aided design, synthetic DNA site-specific recombinases. Using Cre recombinase swapped 126-kb segment the Escherichia coli with 72-kb cassette, thereby effectively eliminating over 54 kb genomic from three non-contiguous regions single recombination event. We observed complete replacement native sequence modified through action no competition homologous recombination. Because versatility high-efficiency Cre-lox system, this method can be used any organism where system is functional as well adapted to use other highly precise systems. Compared present-day iterative approaches engineering, anticipate will greatly speed up creation reduced, modularized optimized genomes integration deletion analyses data, transcriptomics, biology
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