ArticleNucleic acids research2017
Cloning-independent markerless gene editing in Streptococcus sanguinis: novel insights in type IV pilus biology.
Article in Nucleic acids research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 36 citations in OpenAlex.
- A metagenomic study of the gut microbiome in patients with type 2 diabetes mellitus and myocardial infarction.Acta diabetologica · 2026Article
- Expanding the genetic toolbox forApplied and environmental microbiology · 2024Article
- Article
- Structure of a heteropolymeric type 4 pilus from a monoderm bacterium.Nature communications · 2023Article
- Glycerol metabolism supports oral commensal interactions.The ISME journal · 2023Article
- Mechanism of assembly of type 4 filaments: everything you always wanted to know (but were afraid to ask).Microbiology (Reading, England) · 2023Review
- Post-vaccine epidemiology of serotype 3 pneumococci identifies transformation inhibition through prophage-driven alteration of a non-coding RNA.Genome medicine · 2022Article
- Article
- Type IV Pili of Streptococcus sanguinis Contribute to Pathogenesis in Experimental Infective Endocarditis.Microbiology spectrum · 2021Article
- PilB fromProceedings of the National Academy of Sciences of the United States of America · 2021Article
- Advancing Genetic Tools inGenes · 2020Article
- DNA-uptake pili of Vibrio cholerae are required for chitin colonization and capable of kin recognition via sequence-specific self-interaction.Nature microbiology · 2019Article
- Global biochemical and structural analysis of the type IV pilus from the Gram-positive bacteriumThe Journal of biological chemistry · 2019Article
- Gram-Positive Type IV Pili and Competence.Microbiology spectrum · 2019Article
- QstR-dependent regulation of natural competence and type VI secretion in Vibrio cholerae.Nucleic acids research · 2018Article
- Molecular insights into Vibrio cholerae's intra-amoebal host-pathogen interactions.Nature communications · 2018Article
- Streptococcus sanguinis biofilm formation & interaction with oral pathogens.Future microbiology · 2018Review
- Surfaceome and Proteosurfaceome in Parietal Monoderm Bacteria: Focus on Protein Cell-Surface Display.Frontiers in microbiology · 2018Review
- Structure of the competence pilus major pilin ComGC inThe Journal of biological chemistry · 2017Article
- Recombineering inFrontiers in cellular and infection microbiology · 2017Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Streptococcus sanguinis, a naturally competent opportunistic human pathogen, is a Gram-positive workhorse for genomics. It has recently emerged as a model for the study of type IV pili (Tfp)-exceptionally widespread and important prokaryotic filaments. To enhance genetic manipulation of Streptococcus sanguinis, we have developed a cloning-independent methodology, which uses a counterselectable marker and allows sophisticated markerless gene editing in situ. We illustrate the utility of this methodology by answering several questions regarding Tfp biology by (i) deleting single or mutiple genes, (ii) altering specific bases in genes of interest, and (iii) engineering genes to encode proteins with appended affinity tags. We show that (i) the last six genes in the pil locus harbouring all the genes dedicated to Tfp biology play no role in piliation or Tfp-mediated motility, (ii) two highly conserved Asp residues are crucial for enzymatic activity of the prepilin peptidase PilD and (iii) that pilin subunits with a C-terminally appended hexa-histidine (6His) tag are still assembled into functional Tfp. The methodology for genetic manipulation we describe here should be broadly applicable.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.