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

Nature Communications
Wenjun JiangTing F Zhu

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.

References

Apr 25, 1991·Nucleic Acids Research·V Zawadzki, H J Gross
Jan 1, 1971·Folia Microbiologica·Z VanĕkZ Hostálek
Mar 15, 1994·Proceedings of the National Academy of Sciences of the United States of America·W M Barnes
Nov 25, 2005·Nature·Drew Endy
Mar 24, 2007·Science·Rodolphe BarrangouPhilippe Horvath
Mar 1, 2008·Science·Brent C ChristnerDavid C Sands
Apr 14, 2009·Nature Methods·Daniel G GibsonHamilton O Smith
Dec 1, 2009·Cell·Caryn R HaleMichael P Terns
Nov 5, 2010·Nature·Josiane E GarneauSylvain Moineau
Aug 5, 2011·Nucleic Acids Research·Rimantas SapranauskasVirginijus Siksnys
Feb 9, 2012·Metabolic Engineering·Jay D Keasling
Feb 14, 2012·Methods in Molecular Biology·Mamie Z Li, Stephen J Elledge
Feb 22, 2012·Current Opinion in Chemical Biology·Fong T Wong, Chaitan Khosla
May 9, 2012·Nature Biotechnology·Ryan E Cobb, Huimin Zhao
Jun 30, 2012·Science·Martin JinekEmmanuelle Charpentier
Apr 30, 2013·Current Opinion in Chemical Biology·Luisa S GronenbergJames C Liao
May 9, 2013·ACS Synthetic Biology·Rui-Yan WangGuo-Qiang Chen
Jul 23, 2013·Nature Biotechnology·Patrick D HsuFeng Zhang
Sep 21, 2013·ACS Synthetic Biology·François St-PierreKeith E Shearwin
Nov 5, 2013·Journal of Bacteriology·Carol Vargas-BautistaPaul Straight
Nov 22, 2013·Biochemical Society Transactions·Tautvydas KarvelisVirginijus Siksnys
Dec 18, 2013·Science·Tim WangEric S Lander
Jan 22, 2014·Nature Communications·Jong Min KimJin-Soo Kim
Jan 23, 2014·Proceedings of the National Academy of Sciences of the United States of America·Kazuya YamanakaBradley S Moore
Jan 28, 2014·Nature Biotechnology·Yanfang FuJ Keith Joung
Jan 31, 2014·Nature·Samuel H SternbergJennifer A Doudna
Sep 4, 2014·Nature Biotechnology·John G DoenchDavid E Root
Nov 29, 2014·Science·Jennifer A Doudna, Emmanuelle Charpentier

❮ Previous
Next ❯

Citations

Feb 28, 2018·Proceedings of the National Academy of Sciences of the United States of America·Wei TangYihua Chen
Oct 5, 2017·Nucleic Acids Research·Pamela E Bennett-Baker, Jacob L Mueller
Apr 14, 2016·Natural Product Reports·Mingzi M ZhangHuimin Zhao
Dec 23, 2015·Natural Product Reports·Eric J N Helfrich, Jörn Piel
Jan 9, 2019·Natural Product Reports·Liujie HuoRolf Müller
Feb 8, 2019·Biotechnology and Bioengineering·Liping WangYunzi Luo
Feb 28, 2019·Science China. Life Sciences·José Ranz, Bryan Clifton
Jan 30, 2019·Conservation Biology : the Journal of the Society for Conservation Biology·Michael P PhelpsJames E Seeb
Aug 20, 2019·Science China. Life Sciences·Wei Li, Wen-Bing Yin
Nov 18, 2018·Journal of Industrial Microbiology & Biotechnology·Min Xu, Gerard D Wright
May 28, 2019·Frontiers in Genetics·Tuomo MantereAlexander Hoischen
Jun 15, 2019·Natural Product Reports·Jia Jia ZhangBradley S Moore
Jun 27, 2019·Applied Microbiology and Biotechnology·Xuefeng WangWeishan Wang
Aug 15, 2018·Applied Microbiology and Biotechnology·Jia Jia ZhangXiaoyu Tang
Nov 20, 2018·Acta Biochimica Et Biophysica Sinica·Jingman WangGuoping Zhao
Apr 24, 2019·Nature Methods·Carolin A MüllerConrad A Nieduszynski
Dec 15, 2018·Natural Product Reports·Yaojun TongSang Yup Lee
Dec 20, 2019·Journal of Industrial Microbiology & Biotechnology·Ran LiuTiangang Liu
May 3, 2018·FEMS Microbiology Letters·Arianna TocchettiMargherita Sosio
Apr 26, 2019·The CRISPR Journal·Matthew Loose
Dec 7, 2018·Genetics·Behnom FarboudBarbara J Meyer
Jan 31, 2020·Frontiers in Genetics·Lukas F K KudernaFerran Casals
Mar 12, 2019·Engineering in Life Sciences·Qiyuan ZhaoYunzi Luo
Jul 21, 2020·Applied Microbiology and Biotechnology·Jan KormanecDagmar Homerova
Jun 26, 2020·Frontiers in Bioengineering and Biotechnology·Zhenquan LinZihe Liu
Jun 20, 2018·Journal of Industrial Microbiology & Biotechnology·Leonard KatzRichard H Baltz

❮ Previous
Next ❯

Methods Mentioned

BETA
PCR
restriction digest
electrophoresis
in vitro transcription

Software Mentioned

CATCH

Related Concepts

Related Feeds

Biophysics of CRISPR

This feed focuses on broad characteristics of the CRISPR system and the proteins associated with it.

Related Papers

Journal of Molecular Cell Biology
Dongshan YangY Eugene Chen
Methods in Molecular Biology
Rui T L Rodrigues, Travis S Bayer
Journal of Microbiological Methods
Antony T VincentSteve J Charette
© 2021 Meta ULC. All rights reserved