Cas9-Assisted Targeting of CHromosome segments CATCH enables one-step targeted cloning of large gene clusters

• DOI: 10.1038/ncomms9101
• Publication Date: 2015-09-01T10:05:20Z
• Authors: Wenjun Jiang; Xuejin Zhao; Tslil Gabrieli; Chunbo Lou; Yuval Ebenstein; Ting F. Zhu
• Keywords: Cloning (programming); Synthetic Biology

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.