A pooled CRISPRi screen to identify new cell cycle proteins in the opportunistic human pathogen Streptococcus pneumoniae

Informations projet

GetToKnowPneumo

N° de convention de subvention: 837923

DOI

10.3030/837923

Projet clôturé

Date de signature de la CE 21 Mars 2019

Date de début 1 Avril 2019

Date de fin 31 Mars 2021

Financé au titre de

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Coût total

€ 191 149,44

Contribution de l’UE

€ 191 149,44

Coordonné par

UNIVERSITE DE LAUSANNE
Switzerland

Periodic Reporting for period 1 - GetToKnowPneumo (A pooled CRISPRi screen to identify new cell cycle proteins in the opportunistic human pathogen Streptococcus pneumoniae)

Période du rapport: 2019-04-01 au 2021-03-31

Résumé du contexte et des objectifs généraux du projet

Streptococcus pneumoniae is an opportunistic human pathogen that is responsible for diseases such as pneumonia, middle ear infections, sepsis and meningitis and is an important cause of fatal bacterial infections. Moreover, antibiotic resistant S. pneumoniae strains are on the rise, thereby limiting effective treatment options. For these reasons, we set out to gain a better understanding of the cell biology of S. pneumoniae. Such increased knowledge not only has great fundamental scientific value, but could also be exploited in the design and development of novel antibacterial therapies.

Travail effectué depuis le début du projet jusqu’à la fin de la période considérée dans le rapport et principaux résultats atteints jusqu’à présent

Using an innovative, genome-wide, pooled CRISPRi screening approach coupled to sorting of cells that display a phenotype of interest, we have identified genes involved in the regulation of S. pneumoniae cell size, DNA content and cell division, features that are important markers for cell cycle progression. This screen identified several targets that interfere with these processes, including genes of unknown and/or seemingly unrelated function. Our results therefore contribute towards a better understanding of these processes. Moreover, initial tests have shown the potential of some of these hits for the development of novel antibacterial therapies, which will be pursued further in the future.

Results will be disseminated shortly through publication of a scientific manuscript. Moreover, a potential patent application for novel antibacterial therapies is being prepared.

Progrès au-delà de l’état des connaissances et impact potentiel prévu (y compris l’impact socio-économique et les conséquences sociétales plus larges du projet jusqu’à présent)

Although several bacterial genome-wide CRISPRi screens have been performed so far, none of these screen designs included the sorting of cells that display a phenotype of interest. By combining CRISPRi with high-throughput selection of desired phenotypes, we go beyond the state of the art and expand the use of pooled genome-wide CRISPRi screens beyond the investigation of fitness effects, as is current practice.

Moreover, our screen allowed us to identify a role for genes of unknown function in maintenance of S. pneumoniae cell size, DNA content and/or cell division, thereby further increasing our knowledge of these processes and genes.

Finally, we believe to have identified novel drug targets that could form the basis of innovative antibacterial therapies that are effective against S. pneumoniae. Follow-up work will explore these targets further and will hopefully result in the development of novel treatment options.

Overview of our CRISPRi-FACS-seq screen