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CRISPAIR SIGNED

Study of the interplay between CRISPR interference and DNA repair pathways towards the development of novel CRISPR tools

Total Cost €

EC-Contrib. €

Partnership

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Outcomes and
results

CRISPAIR project word cloud

Explore the words cloud of the CRISPAIR project. It provides you a very rough idea of what is the project "CRISPAIR" about.

interaction combinations combination nucleases processed critical always knockouts biotechnological edit tools kill light eukaryotic cleavage chromosome crispr capture automation bacterial elusive bacteria introduce advantage repaired archaea immune perform genetic first compete genetics nhej loci data coli fascinating generate rna preliminary cell guided survive interplay dna genome cas gene cells degrade breaks invading consortiums mutations immunity multiplexed employed popular tool wise repair biology cas9 indicates strategies bioinformatics action pieces shed time organism knockout introduction libraries gained engineering screens pair networks genomes adaptive evolution decipher

Project "CRISPAIR" data sheet

The following table provides information about the project.

Coordinator	INSTITUT PASTEUR Organization address address: RUE DU DOCTEUR ROUX 25-28 city: PARIS CEDEX 15 postcode: 75724 website: http://www.pasteur.fr contact info title: n.a. name: n.a. surname: n.a. function: n.a. email: n.a. telephone: n.a. fax: n.a.
Coordinator Country	France [FR]
Total cost	1˙499˙763 €
EC max contribution	1˙499˙763 € (100%)
Programme	1. H2020-EU.1.1. (EXCELLENT SCIENCE - European Research Council (ERC))
Code Call	ERC-2015-STG
Funding Scheme	ERC-STG
Starting year	2016
Duration (year-month-day)	from 2016-03-01 to 2021-11-30

Partnership

Take a look of project's partnership.

#	participants	country	role	EC contrib. [€]
1	INSTITUT PASTEUR INSTITUT PASTEUR Organization address address: RUE DU DOCTEUR ROUX 25-28 city: PARIS CEDEX 15 postcode: 75724 website: http://www.pasteur.fr contact info title: n.a. name: n.a. surname: n.a. function: n.a. email: n.a. telephone: n.a. fax: n.a.	FR (PARIS CEDEX 15)	coordinator	1˙499˙763.00

Map

Project objective

CRISPR-Cas loci are the adaptive immune system of archaea and bacteria. They can capture pieces of invading DNA and use this information to degrade target DNA through the action of RNA-guided nucleases. The consequences of DNA cleavage by Cas nucleases, i.e. how breaks are processed and whether they can be repaired, remains to be investigated. A better understanding of the interplay between DNA repair and CRISPR-Cas is critical both to shed light on the evolution and biology of these fascinating systems and for the development of biotechnological tools based on Cas nucleases. CRISPR systems have indeed become a popular tool to edit Eukaryotic genomes. The strategies employed take advantage of different DNA repair pathways to introduce mutations upon DNA cleavage. In bacteria however, the introduction of breaks by Cas nucleases in the chromosome has been described to kill the cell. Preliminary data indicates that this might not always be the case and that some DNA repair pathways could compete with CRISPR immunity allowing cells to survive. Using a combination of bioinformatics and genetics approaches we will investigate the interplay between CRISPR and DNA repair in bacteria with a particular focus on the widely used CRISPR-Cas9 system. The knowledge gained from this study will then help us develop novel tools for bacterial genome engineering. In particular we will introduce a NHEJ pathway in E.coli making it possible to perform CRISPR knockout screens. Finally using CRISPR libraries and multiplexed targeting, we will generate for the first time all combinations of pair-wise gene knockouts in an organism, a task that for now remains elusive, even for large consortiums and with the use of automation. This will enable to decipher genome-scale genetic interaction networks, an important step for our understanding of bacteria as a system.

Publications

List of publications.
year	authors and title	journal	last update
2017	David Bikard, Rodolphe Barrangou Using CRISPR-Cas systems as antimicrobials published pages: 155-160, ISSN: 1369-5274, DOI: 10.1016/j.mib.2017.08.005	Current Opinion in Microbiology 37	2020-01-21
2018	FranÃ§ois Rousset, Lun Cui, Elise Siouve, Christophe Becavin, Florence Depardieu, David Bikard Genome-wide CRISPR-dCas9 screens in E. coli identify essential genes and phage host factors published pages: e1007749, ISSN: 1553-7404, DOI: 10.1371/journal.pgen.1007749	PLOS Genetics 14/11	2020-01-21
2018	Lun Cui, Antoine Vigouroux, FranÃ§ois Rousset, Hugo Varet, Varun Khanna, David Bikard A CRISPRi screen in E. coli reveals sequence-specific toxicity of dCas9 published pages: , ISSN: 2041-1723, DOI: 10.1038/s41467-018-04209-5	Nature Communications 9/1	2020-01-21
2017	Aude Bernheim, Alicia Calvo-VillamaÃ±Ã¡n, Clovis Basier, Lun Cui, Eduardo P. C. Rocha, Marie Touchon, David Bikard Inhibition of NHEJ repair by type II-A CRISPR-Cas systems in bacteria published pages: , ISSN: 2041-1723, DOI: 10.1038/s41467-017-02350-1	Nature Communications 8/1	2020-01-21
2018	Antoine Vigouroux, Enno Oldewurtel, Lun Cui, David Bikard, Sven van Teeffelen Tuning dCas9\'s ability to block transcription enables robust, noiseless knockdown of bacterial genes published pages: e7899, ISSN: 1744-4292, DOI: 10.15252/msb.20177899	Molecular Systems Biology 14/3	2020-01-21

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