Data on the link between genomic integration of IS1548 and lineage of the strain obtained by bioinformatic analyses of sequenced genomes of Streptococcus agalactiae available at the National Center for Biotechnology Information database

Khazaal, Sarah; Safadi, Rim Al; Osman, Dani; Hiron, Aurélia; Gilot, Philippe

doi:10.1016/j.dib.2019.105066

Cited by 2 publications

(1 citation statement)

References 4 publications

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“…The long-read genome-sequenced NV1 strain was compared to the published 5448 reference genome which was generated by illumina short read sequencing (25), revealing a large inversion of 1,475,033 base pairs. This structural variant is flanked by two ISAs1-like element IS1548 family transposase genes, which typically harbor terminal inverted repeats (26). The two transposase genes are oriented in opposing directions and exhibit two single nucleotide polymorphisms.…”

Section: Resultsmentioning

confidence: 99%

An efficientin vivo-inducible CRISPR interference system for group AStreptococcusgenetic analysis and pathogenesis studies

Bjånes,

Stream,

Janssen

et al. 2024

Preprint

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While genome-wide transposon mutagenesis screens have identified numerous essential genes in the significant human pathogenStreptococcus pyogenes(group AStreptococcusor GAS), many of their functions remain elusive. This knowledge gap is attributed in part to the limited molecular toolbox for controlling GAS gene expression and the bacterium’s poor genetic transformability. CRISPR interference (CRISPRi), using catalytically inactive GAS Cas9 (dCas9), is a powerful approach to specifically repress gene expression in both bacteria and eukaryotes, but ironically has never been harnessed for controlled gene expression in GAS. In this study, we present a highly transformable and fully virulent serotype M1T1 GAS strain and introduce a doxycycline-inducible CRISPRi system for efficient repression of bacterial gene expression. We demonstrate highly efficient, oligo-based sgRNA cloning directly to GAS, enabling the construction of a gene knockdown strain in just two days, in contrast to the several weeks typically required. The system is shown to be titratable and functional bothin vitroandin vivousing a murine model of GAS infection. Furthermore, we provide directin vivoevidence that the expression of the conserved cell division geneftsZis essential for GAS virulence, highlighting its promise as a target for emerging FtsZ-inhibitors. Finally, we introduce SpyBrowse (https://veeninglab.com/SpyBrowse), a comprehensive and user-friendly online resource for visually inspecting and exploring GAS genetic features. The tools and methodologies described in this work are poised to facilitate fundamental research in GAS, contribute to vaccine development, and aid in the discovery of antibiotic targets.Significance statementWhile GAS remains a predominant cause of bacterial infections worldwide, there are limited genetic tools available to study its basic cell biology. Here, we bridge this gap by creating a highly transformable, fully virulent M1T1 GAS strain. In addition, we established a tight and titratable doxycycline-inducible system and developed CRISPR interference for controlled gene expression in GAS. We show that CRISPRi is functionalin vivoin a mouse infection model. Additionally, we present SpyBrowse, an intuitive and accessible genome browser (https://veeninglab.com/SpyBrowse). Overall, this work overcomes significant technical challenges of working with GAS, and together with SpyBrowse, represents a valuable resource for researchers in the GAS field.

show abstract

Section: Resultsmentioning

confidence: 99%

An efficientin vivo-inducible CRISPR interference system for group AStreptococcusgenetic analysis and pathogenesis studies

Bjånes,

Stream,

Janssen

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

An efficient in vivo -inducible CRISPR interference system for group A Streptococcus genetic analysis and pathogenesis studies

Bjånes,

Stream,

Janssen

et al. 2024

mBio

View full text Add to dashboard Cite

While genome-wide transposon mutagenesis screens have identified numerous essential genes in the significant human pathogen Streptococcus pyogenes (group A Streptococcus or GAS), many of their functions remain elusive. This knowledge gap is attributed in part to the limited molecular toolbox for controlling GAS gene expression and the bacterium’s poor genetic transformability. CRISPR interference (CRISPRi), using catalytically inactive GAS Cas9 (dCas9), is a powerful approach to specifically repress gene expression in both bacteria and eukaryotes, but ironically, it has never been harnessed for controlled gene expression in GAS. In this study, we present a highly transformable and fully virulent serotype M1T1 GAS strain and introduce a doxycycline-inducible CRISPRi system for efficient repression of bacterial gene expression. We demonstrate highly efficient, oligo-based single guide RNA cloning directly to GAS, enabling the construction of a gene knockdown strain in just 2 days, in contrast to the several weeks typically required. The system is shown to be titratable and functional both in vitro and in vivo using a murine model of GAS infection. Furthermore, we provide direct in vivo evidence that the expression of the conserved cell division gene ftsZ is essential for GAS virulence, highlighting its promise as a target for emerging FtsZ inhibitors. Finally, we introduce SpyBrowse ( https://veeninglab.com/SpyBrowse ), a comprehensive and user-friendly online resource for visually inspecting and exploring GAS genetic features. The tools and methodologies described in this work are poised to facilitate fundamental research in GAS, contribute to vaccine development, and aid in the discovery of antibiotic targets. IMPORTANCE While group A Streptococcus (GAS) remains a predominant cause of bacterial infections worldwide, there are limited genetic tools available to study its basic cell biology. Here, we bridge this gap by creating a highly transformable, fully virulent M1T1 GAS strain. In addition, we established a tight and titratable doxycycline-inducible system and developed CRISPR interference (CRISPRi) for controlled gene expression in GAS. We show that CRISPRi is functional in vivo in a mouse infection model. Additionally, we present SpyBrowse, an intuitive and accessible genome browser ( https://veeninglab.com/SpyBrowse ). Overall, this work overcomes significant technical challenges of working with GAS and, together with SpyBrowse, represents a valuable resource for researchers in the GAS field.

show abstract

Data on the link between genomic integration of IS1548 and lineage of the strain obtained by bioinformatic analyses of sequenced genomes of Streptococcus agalactiae available at the National Center for Biotechnology Information database

Cited by 2 publications

References 4 publications

An efficientin vivo-inducible CRISPR interference system for group AStreptococcusgenetic analysis and pathogenesis studies

An efficientin vivo-inducible CRISPR interference system for group AStreptococcusgenetic analysis and pathogenesis studies

An efficient in vivo -inducible CRISPR interference system for group A Streptococcus genetic analysis and pathogenesis studies

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