Efficient genome editing for Pseudomonas aeruginosa using CRISPR-Cas12a.

Gene
Zhanglin LinYanrui Ye

Abstract

The CRISPR-Cas12a system has been demonstrated as an attractive tool for bacterial genome engineering. In particular, FnCas12a recognizes protospacer-adjacent motif (PAM) sites with medium or low GC content, which complements the Cas9-based systems. Here we explored Francisella novicida Cas12a (FnCas12a) for genome editing in Pseudomonas aeruginosa. By using a two-plasmid system expressing the constitutive FnCas12a nuclease, the inducible λRed recombinase, a CRISPR RNA (crRNA), we achieved gene deletion, insertion and replacement with high efficiency (in most cases > 75%), including the deletion of large DNA fragments up to 15 kb and the serial deletion of duplicate gene clusters. This work should provide a useful and complementary addition to the genome engineering toolbox for the study of P. aeruginosa biology and physiology.

References

Jan 1, 1981·Molecular & General Genetics : MGG·D M StalkerD R Helinski
Nov 26, 1999·Proceedings of the National Academy of Sciences of the United States of America·M WhiteleyE P Greenberg
Feb 6, 2008·BMC Molecular Biology·Biliana Lesic, Laurence G Rahme
Nov 4, 2008·Nucleic Acids Research·Geoffrey L WinsorFiona S L Brinkman
Jan 31, 2009·Nature Protocols·Shyam K SharanDonald L Court
Apr 14, 2009·Nature Methods·Daniel G GibsonHamilton O Smith
Aug 14, 2010·Journal of Medical Microbiology·Haihua LiangKangmin Duan
Jun 15, 2011·Trends in Microbiology·Elena B M BreidensteinRobert E W Hancock
Jun 21, 2011·Journal of Cystic Fibrosis : Official Journal of the European Cystic Fibrosis Society·Pieter DeschaghtMario Vaneechoutte
Jun 30, 2012·Science·Martin JinekEmmanuelle Charpentier
Aug 28, 2012·Applied Microbiology and Biotechnology·Hui ShiKunlun Huang
Jan 5, 2013·Science·Prashant MaliGeorge M Church
Jan 31, 2013·Nature Biotechnology·Wenyan JiangLuciano A Marraffini
Jul 31, 2014·Nucleic Acids Research·Jee-Hwan Oh, Jan-Peter van Pijkeren
Feb 1, 2015·Applied and Environmental Microbiology·Yu JiangSheng Yang
May 15, 2015·Journal of Virology·Diana P PiresJoana Azeredo
Oct 23, 2015·Nature Protocols·Laura R HmeloJoe J Harrison
Apr 5, 2016·Molecular Cell·Ryan T LeenayChase L Beisel
Jun 9, 2016·BioMed Research International·Sirijan Santajit, Nitaya Indrawattana
Jul 28, 2016·Current Microbiology·Tsiry Rasamiravaka, Mondher El Jaziri
Oct 30, 2016·Applied and Environmental Microbiology·Indira Cruz-PlancarteGabriel Guarneros
Dec 22, 2016·Scientific Reports·Justin Ungerer, Himadri B Pakrasi
Feb 22, 2017·Journal of the American Chemical Society·Weizhong ChenQuanjiang Ji
May 5, 2017·Nature Communications·Yu JiangSheng Yang
Jun 14, 2017·Cell Discovery·Xiaochun ZhangJin Wang
Aug 2, 2017·Nature Medicine·David A Scott, Feng Zhang
Sep 21, 2017·BMC Microbiology·Weiliang Huang, Angela Wilks
Oct 24, 2017·Biotechnology Journal·Tomás AparicioEsteban Martínez-García
Apr 6, 2018·International Journal of Molecular Sciences·Suhyung ChoByung-Kwan Cho
May 24, 2018·Wiley Interdisciplinary Reviews. RNA·Daan C Swarts, Martin Jinek

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