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

Wenjun Jiang, Xuejin Zhao, Tslil Gabrieli, Chunbo Lou (), Yuval Ebenstein () and Ting F. Zhu ()
Additional contact information
Wenjun Jiang: School of Life Sciences, Center for Synthetic and Systems Biology, MOE Key Laboratory of Bioinformatics, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University
Xuejin Zhao: CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences
Tslil Gabrieli: Raymond and Beverly Sackler Faculty of Exact Sciences, School of Chemistry, Tel Aviv University
Chunbo Lou: CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences
Yuval Ebenstein: Raymond and Beverly Sackler Faculty of Exact Sciences, School of Chemistry, Tel Aviv University
Ting F. Zhu: School of Life Sciences, Center for Synthetic and Systems Biology, MOE Key Laboratory of Bioinformatics, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University

Nature Communications, 2015, vol. 6, issue 1, 1-8

Abstract: Abstract The cloning of long DNA segments, especially those containing large gene clusters, is of particular importance to synthetic and chemical biology efforts for engineering organisms. While cloning has been a defining tool in molecular biology, the cloning of long genome segments has been challenging. Here we describe a technique that allows the targeted cloning of near-arbitrary, long bacterial genomic sequences of up to 100 kb to be accomplished in a single step. The target genome segment is excised from bacterial chromosomes in vitro by the RNA-guided Cas9 nuclease at two designated loci, and ligated to the cloning vector by Gibson assembly. This technique can be an effective molecular tool for the targeted cloning of large gene clusters that are often expensive to synthesize by gene synthesis or difficult to obtain directly by traditional PCR and restriction-enzyme-based methods.

Date: 2015
References: Add references at CitEc
Citations: View citations in EconPapers (6)

Downloads: (external link)
https://www.nature.com/articles/ncomms9101 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms9101

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/ncomms9101

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().