Cas9-Assisted Targeting of CHromosome segments CATCH enables one-step targeted cloning of large gene clusters
Cloning (programming)
Synthetic Biology
DOI:
10.1038/ncomms9101
Publication Date:
2015-09-01T10:05:20Z
AUTHORS (6)
ABSTRACT
Abstract The cloning of long DNA segments, especially those containing large gene clusters, is particular importance to synthetic and chemical biology efforts for engineering organisms. While has been a defining tool in molecular biology, the genome segments challenging. Here we describe technique that allows targeted near-arbitrary, bacterial genomic sequences up 100 kb be accomplished single step. target segment excised from chromosomes vitro by RNA-guided Cas9 nuclease at two designated loci, ligated vector Gibson assembly. This can an effective clusters are often expensive synthesize synthesis or difficult obtain directly traditional PCR restriction-enzyme-based methods.
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