CRISPR-FRT targets shared sites in a knock-out collection for off-the-shelf genome editing

Toon Swings; David C Marciano; Benu Atri; Rachel E Bosserman; Chen Wang; Marlies Leysen; Camille Bonte; Thomas Schalck; Ian Furey; Bram Van den Bergh; Natalie Verstraeten; Peter J Christie; Christophe Herman; Olivier Lichtarge; Jan Michiels

doi:10.1038/s41467-018-04651-5

CRISPR-FRT targets shared sites in a knock-out collection for off-the-shelf genome editing

Nat Commun. 2018 Jun 8;9(1):2231. doi: 10.1038/s41467-018-04651-5.

Authors

Toon Swings^{1

2}, David C Marciano³, Benu Atri⁴, Rachel E Bosserman⁵, Chen Wang³, Marlies Leysen¹, Camille Bonte¹, Thomas Schalck^{1

2}, Ian Furey⁶, Bram Van den Bergh^{1

2}, Natalie Verstraeten^{1

2}, Peter J Christie⁵, Christophe Herman³, Olivier Lichtarge^{7

8

9

10}, Jan Michiels^{11

12}

Affiliations

¹ Centre of Microbial and Plant Genetics, KU Leuven - University of Leuven, Kasteelpark Arenberg 20, 3001, Leuven, Belgium.
² Center for Microbiology, VIB, Kasteelpark Arenberg 20, 3001, Leuven, Belgium.
³ Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, 77030, USA.
⁴ Quantitative and Computational Biosciences, Baylor College of Medicine, Houston, TX, 77030, USA.
⁵ Department of Microbiology and Molecular Genetics, McGovern Medical School, Houston, TX, 77030, USA.
⁶ Department of Pharmacology, Baylor College of Medicine, Houston, TX, 77030, USA.
⁷ Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, 77030, USA. lichtarge@bcm.edu.
⁸ Quantitative and Computational Biosciences, Baylor College of Medicine, Houston, TX, 77030, USA. lichtarge@bcm.edu.
⁹ Department of Pharmacology, Baylor College of Medicine, Houston, TX, 77030, USA. lichtarge@bcm.edu.
¹⁰ Computational and Integrative Biomedical Research Center, Baylor College of Medicine, Houston, TX, 77030, USA. lichtarge@bcm.edu.
¹¹ Centre of Microbial and Plant Genetics, KU Leuven - University of Leuven, Kasteelpark Arenberg 20, 3001, Leuven, Belgium. jan.michiels@kuleuven.vib.be.
¹² Center for Microbiology, VIB, Kasteelpark Arenberg 20, 3001, Leuven, Belgium. jan.michiels@kuleuven.vib.be.

Abstract

CRISPR advances genome engineering by directing endonuclease sequence specificity with a guide RNA molecule (gRNA). For precisely targeting a gene for modification, each genetic construct requires a unique gRNA. By generating a gRNA against the flippase recognition target (FRT) site, a common genetic element shared by multiple genetic collections, CRISPR-FRT circumvents this design constraint to provide a broad platform for fast, scarless, off-the-shelf genome engineering.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Binding Sites / genetics
CRISPR-Cas Systems*
DNA Nucleotidyltransferases / genetics
DNA Nucleotidyltransferases / metabolism*
DNA, Bacterial / genetics
DNA, Bacterial / metabolism
Escherichia coli / genetics
Escherichia coli / metabolism
Gene Editing / methods*
Gene Knockout Techniques
Genome, Bacterial / genetics
Models, Genetic
Mutation
RNA, Guide / genetics
RNA, Guide / metabolism*

Substances

DNA, Bacterial
RNA, Guide
DNA Nucleotidyltransferases
Site-specific recombinase

Abstract

Publication types

MeSH terms

Substances

Grant support