Genetic manipulation of Staphylococci—breaking through the barrier

Most strains of Staphylococcus aureus and Staphylococcus epidermidis possess a strong restriction barrier that hinders exchange of DNA. Recently, major advances have been made in identifying and characterizing the restriction-modification (RM) systems involved. In particular a novel type IV restriction enzyme that recognizes cytosine methylated DNA has been shown to be the major barrier to transfer of plasmid DNA from Escherichia coli into S. aureus and S. epidermidis. While the conserved type I RM system provides a further barrier. Here we review the recent advances in understanding of restriction systems in staphylococci and highlight how this has been exploited to improve our ability to manipulate genetically previously untransformable strains.

• Publication year

  2012

• Venue

  Front. Cell. Inf. Microbio.

• Publication date

  2012-04-12

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.3389/fcimb.2012.00049PMID 22919640PMCID 3417578
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Genetic Manipulation of Staphylococcus aureus

  2014cited by this paper
• Genome Sequence of Staphylococcus aureus Strain 11819-97, an ST80-IV European Community-Acquired Methicillin-Resistant Isolate

  2012cited by this paper
• A single copy integration vector that integrates at an engineered site on the Staphylococcus aureus chromosome

  2012cited by this paper
• A single copy integration vector that integrates at an engineered site on the Staphylococcus aureus chromosome

  2012cited by this paper
• High efficiency recombineering in lactic acid bacteria

  2012cited by this paper
• Transforming the Untransformable: Application of Direct Transformation To Manipulate Genetically Staphylococcus aureus and Staphylococcus epidermidis

  2012cited by this paper
• Vectors for improved Tet repressor-dependent gradual gene induction or silencing in Staphylococcus aureus.

  2011cited by this paper
• A simple method of markerless gene deletion in Staphylococcus aureus.

  2011cited by this paper
• Plasmid artificial modification: a novel method for efficient DNA transfer into bacteria.

  2011cited by this paper
• Engineering multiple genomic deletions in Gram-negative bacteria: analysis of the multi-resistant antibiotic profile of Pseudomonas putida KT2440.

  2011cited by this paper
• Rapid and Highly Efficient Method for Scarless Mutagenesis within the Salmonella enterica Chromosome

  2011cited by this paper
• Mutation of tagO reveals an essential role for wall teichoic acids in Staphylococcus epidermidis biofilm development.

  2011cited by this paper
• Recycling of protein subunits during DNA translocation and cleavage by Type I restriction-modification enzymes

  2011cited by this paper
• A type IV modification-dependent restriction enzyme SauUSI from Staphylococcus aureus subsp. aureus USA300

  2011cited by this paper
• Actin Polymerization Drives Septation of Listeria monocytogenes namA Hydrolase Mutants, Demonstrating Host Correction of a Bacterial Defect

  2011cited by this paper
• Whole-Genome Sequencing of Staphylococcus aureus Strain RN4220, a Key Laboratory Strain Used in Virulence Research, Identifies Mutations That Affect Not Only Virulence Factors but Also the Fitness of the Strain

  2011cited by this paper
• Noninvasive in vivo imaging to evaluate immune responses and antimicrobial therapy against Staphylococcus aureus and USA300 MRSA skin infections.

  2011cited by this paper
• Mobile genetic elements of Staphylococcus aureus

  2010cited by this paper
• Oligonucleotide recombination in Gram‐negative bacteria

  2010cited by this paper
• Aureusimines in Staphylococcus aureus Are Not Involved in Virulence

  2010cited by this paper
• Genomic variation and evolution of Staphylococcus aureus.

  2010influential reference
• Taking phage integration to the next level as a genetic tool for mycobacteria.

  2010cited by this paper
• Polymorphonuclear leukocytes mediate Staphylococcus aureus Panton-Valentine leukocidin-induced lung inflammation and injury

  2010cited by this paper
• A type III-like restriction endonuclease functions as a major barrier to horizontal gene transfer in clinical Staphylococcus aureus strains

  2010influential reference
• Conservation of Genomic Localization and Sequence Content of Sau3AI-Like Restriction-Modification Gene Cassettes among Listeria monocytogenes Epidemic Clone I and Selected Strains of Serotype 1/2a

  2010cited by this paper
• Repair of Global Regulators in Staphylococcus aureus 8325 and Comparative Analysis with Other Clinical Isolates

  2010cited by this paper
• Staphylococcus aureus SH1000 and 8325‐4: comparative genome sequences of key laboratory strains in staphylococcal research

  2010cited by this paper
• PFGE diversity within the methicillin-resistant Staphylococcus aureus clonal lineage ST398

  2010cited by this paper
• Redox sensing by a Rex-family repressor is involved in the regulation of anaerobic gene expression in Staphylococcus aureus

  2010cited by this paper
• Production of the Bsa Lantibiotic by Community-Acquired Staphylococcus aureus Strains

  2009cited by this paper
• An improved tetracycline-inducible expression vector for Staphylococcus aureus.

  2009cited by this paper
• Regulation of the mazEF Toxin-Antitoxin Module in Staphylococcus aureus and Its Impact on sigB Expression

  2009cited by this paper
• Overcoming the restriction barrier to plasmid transformation and targeted mutagenesis in Bifidobacterium breve UCC2003

  2009cited by this paper
• Inactivation of the SauI Type I Restriction-Modification System Is Not Sufficient To Generate Staphylococcus aureus Strains Capable of Efficiently Accepting Foreign DNA

  2009influential reference
• A system for the construction of targeted unmarked gene deletions in the genus Burkholderia.

  2008cited by this paper
• Efficient point mutagenesis in mycobacteria using single‐stranded DNA recombineering: characterization of antimycobacterial drug targets

  2008cited by this paper
• Improvement of bacterial transformation efficiency using plasmid artificial modification

  2008cited by this paper
• Vancomycin-Resistant Staphylococcus aureus Isolates Associated with Inc18-Like vanA Plasmids in Michigan

  2007cited by this paper
• YycH and YycI Interact To Regulate the Essential YycFG Two-Component System in Bacillus subtilis

  2007cited by this paper
• Optimization of electroporation‐mediated transformation: Staphylococcus carnosus as model organism

  2007cited by this paper
• Genome Sequence of Staphylococcus aureus Strain Newman and Comparative Analysis of Staphylococcal Genomes: Polymorphism and Evolution of Two Major Pathogenicity Islands

  2007influential reference
• Rapid determination of hospital-acquired meticillin-resistant Staphylococcus aureus lineages.

  2007cited by this paper
• Improved single-copy integration vectors for Staphylococcus aureus.

  2007cited by this paper
• Staphylococcus aureus Strains That are Hypersusceptible to Resistance Gene Transfer from Enterococci

  2007cited by this paper
• Harnessing homologous recombination in vitro to generate recombinant DNA via SLIC

  2007cited by this paper
• Allelic replacement in Staphylococcus aureus with inducible counter-selection.

  2006influential reference
• Host defenses against Staphylococcus aureus infection require recognition of bacterial lipoproteins

  2006cited by this paper
• N6-methyl-adenine: an epigenetic signal for DNA–protein interactions

  2006cited by this paper
• Routine Markerless Gene Replacement in Bacillus anthracis

  2006cited by this paper
• Sau1: a Novel Lineage-Specific Type I Restriction-Modification System That Blocks Horizontal Gene Transfer into Staphylococcus aureus and between S. aureus Isolates of Different Lineages

  2006influential reference
• Insights into Mechanisms Used by Staphylococcus aureus to Avoid Destruction by Human Neutrophils

  2005influential reference
• A dichotomous epigenetic mechanism governs expression of the LlaJI restriction/modification system

  2005cited by this paper
• Insights on Evolution of Virulence and Resistance from the Complete Genome Analysis of an Early Methicillin-Resistant Staphylococcus aureus Strain and a Biofilm-Producing Methicillin-Resistant Staphylococcus epidermidis Strain

  2005influential reference
• Insights into Mechanisms Used by Staphylococcus aureus to Avoid Destruction by Human Neutrophils1

  2005cited by this paper
• IL-13受体α2降低血吸虫病肉芽肿的炎症反应并延长宿主存活时间[英]/Mentink-Kane MM,Cheever AW，Thompson RW，et al//Proc Natl Acad Sci U S A

  2005cited by this paper
• New Vector for Efficient Allelic Replacement in Naturally Nontransformable, Low-GC-Content, Gram-Positive Bacteria

  2004cited by this paper
• A divIVA null mutant of Staphylococcus aureus undergoes normal cell division.

  2004cited by this paper
• Novel Cassette-Based Shuttle Vector System for Gram-Positive Bacteria

  2004influential reference
• Construction, Characterization, and Use of Two Listeria monocytogenes Site-Specific Phage Integration Vectors

  2003cited by this paper
• Deletion of the Gene Encoding p60 in Listeria monocytogenes Leads to Abnormal Cell Division and Loss of Actin-Based Motility

  2003cited by this paper
• A new staphylococcal sigma factor in the conserved gene cassette: functional significance and implication for the evolutionary processes

  2003cited by this paper
• Construction, Characterization, and Use of Two Listeria monocytogenes Site-Specific Phage Integration Vectors

  2002cited by this paper
• Conditional-Replication, Integration, Excision, and Retrieval Plasmid-Host Systems for Gene Structure-Function Studies of Bacteria

  2001cited by this paper
• Evaluation of a Tetracycline-Inducible Promoter inStaphylococcus aureus In Vitro and In Vivo and Its Application in Demonstrating the Role of sigB in Microcolony Formation

  2001cited by this paper
• Type I Restriction Systems: Sophisticated Molecular Machines (a Legacy of Bertani and Weigle)

  2000influential reference
• Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus.

  2000cited by this paper
• Regulated gene expression in Staphylococcus aureus for identifying conditional lethal phenotypes and antibiotic mode of action.

  2000cited by this paper
• Markerless gene replacement in Escherichia coli stimulated by a double-strand break in the chromosome.

  1999cited by this paper
• Site-specific integration of bacteriophage VWB genome into Streptomyces venezuelae and construction of a VWB-based integrative vector.

  1998cited by this paper
• Inducible production and cellular location of the epidermin biosynthetic enzyme EpiB using an improved staphylococcal expression system.

  1996cited by this paper
• Characterization of Tn917 insertion mutants of Staphylococcus epidermidis affected in biofilm formation

  1996influential reference
• A general system for generating unlabelled gene replacements in bacterial chromosomes

  1996cited by this paper
• Electrotransformation of Staphylococci.

  1995cited by this paper
• Induction of homologous recombination in mammalian chromosomes by using the I-SceI system of Saccharomyces cerevisiae

  1995cited by this paper
• Adhesion properties of mutants of Staphylococcus aureus defective in fibronectin‐binding proteins and studies on the expression of fnb genes

  1995cited by this paper
• The dam and dcm strains of Escherichia coli--a review.

  1994cited by this paper
• High-efficiency gene inactivation and replacement system for gram-positive bacteria

  1993cited by this paper
• Improved method for electroporation of Staphylococcus aureus.

  1992cited by this paper
• Construction of single-copy integration vectors for Staphylococcus aureus.

  1991cited by this paper
• A novel activity in Escherichia coli K-12 that directs restriction of DNA modified at CG dinucleotides

  1991cited by this paper
• Repression and catabolite repression of the lactose operon of Staphylococcus aureus

  1990cited by this paper
• High-frequency transformation ofStaphylococcus aureus by electroporation

  1990cited by this paper
• Transformation of Staphylococcus epidermidis and other staphylococcal species with plasmid DNA by electroporation.

  1990cited by this paper
• Molecular Koch's postulates applied to microbial pathogenicity.

  1988influential reference
• Expression of the cloned toxic shock syndrome toxin 1 gene (tst) in vivo with a rabbit uterine model

  1985cited by this paper
• Mechanism of bacteriophage conversion of lipase activity in Staphylococcus aureus

  1985cited by this paper
• The toxic shock syndrome exotoxin structural gene is not detectably transmitted by a prophage

  1983cited by this paper
• Replication and incompatibility properties of plasmid pE194 in Bacillus subtilis

  1982cited by this paper
• Restriction-deficient mutants of Staphylococcus aureus.

  1977cited by this paper
• Occurrence of a class II restriction endonuclease in Staphylococcus aureus

  1977cited by this paper
• Two restriction and modification systems in Staphylococcus aureus NCTC8325.

  1976cited by this paper
• The effect of heat on the ability of a host strain to support the growth of a Staphylococcus phage.

  1963cited by this paper
• Variations in a related series of staphylococcal bacteriophages.

  1956cited by this paper

• Pan-genome analysis of methicillin-resistant Staphylococcus aureus PSU20 from a hospital in Thailand reveals insights into virulence, antibiotic resistance, and genetic diversity.

  2026cites this paper
• Development of a Tool for High‐Efficiency, Markerless and Iterative Genome Editing in Shouchella clausii

  2026cites this paper
• Whole-genome sequencing to explore genomic profile, pathogenicity, virulence motifs, and pangenome of clinical MRSA isolates

  2025cites this paper
• From prebiotics to engineered microbes: microbe-inspired therapies for atopic dermatitis.

  2025cites this paper
• Synthetically designed anti-defense proteins overcome barriers to bacterial transformation and phage infection

  2025cites this paper
• Development of a Pre-Modification Strategy to Overcome Restriction–Modification Barriers and Enhance Genetic Engineering in Lactococcus lactis for Nisin Biosynthesis

  2025cites this paper
• The Staphylococcus aureus esterase FmtA is essential for wall teichoic acid D-alanylation

  2025cites this paper
• The potential role of Staphylococcal enterotoxin C and TSST-1 in the infection of bovine mammary epithelial cells.

  2025cites this paper
• Eco-engineered remediation: Microbial and rhizosphere-based strategies for heavy metal detoxification

  2025cites this paper
• Emergence of transferable daptomycin resistance in Gram-positive bacteria

  2025cites this paper
• Construction of an efficient electroporation transformation system promotes the application of Targetron in wild-type Paenibacillus elgii 219

  2025cites this paper
• Comprehensive genomic analysis of antibiotic resistance plasmids in animal-associated Staphylococcus aureus in France

  2025cites this paper
• Novel Variants of SCCmec Type IX Identified in Clonal Complex 398 Livestock-Associated Methicillin-Resistant Staphylococcus aureus from Pork Production Systems in Korea

  2025cites this paper
• Use of CRISPR interference for efficient and rapid gene inactivation in Fusobacterium nucleatum

  2024cites this paper
• Antimicrobial Resistance in Coagulase Negative Staphylococci: Genome Analysis and Role of Horizontal Gene Transfer.

  2024cites this paper
• Altered genomic methylation promotes Staphylococcus aureus persistence in hospital environment

  2024cites this paper
• Optimizing phage-based mutant recovery and minimizing heat effect in the construction of transposon libraries in Staphylococcus aureus

  2024cites this paper
• Deletion of codY impairs Staphylococcus epidermidis biofilm formation, generation of viable but non-culturable cells and stimulates cytokine production in human macrophages.

  2024cites this paper
• Occurrence of cfr-Positive Linezolid-Susceptible Staphylococcus aureus and Non-aureus Staphylococcal Isolates from Pig Farms

  2023cites this paper
• Pyochelin biotransformation by Staphylococcus aureus shapes bacterial competition with Pseudomonas aeruginosa in polymicrobial infections

  2023cites this paper
• The Influence of Deletion in the Non-Catalytic Domain of GdpP Mediated by Genome Editing with CRISPR/Cas9 on the Phenotype of Staphylococcus aureus

  2023cites this paper
• Use of CRISPR interference for efficient and rapid gene inactivation in Fusobacterium nucleatum

  2023cites this paper
• The Influence of a Deletion in the Noncatalytic Domain of GdpP Mediated by Genome Editing with CRISPR/Cas9 on the Phenotype of Staphylococcus aureus

  2023cites this paper
• Efficient Genome Editing in Most Staphylococcus aureus by Using the Restriction-Modification System Silent CRISPR-Cas9 Toolkit.

  2023cites this paper
• Staphylococcus epidermidis activates keratinocyte cytokine expression and promotes skin inflammation through the production of phenol-soluble modulins

  2023cites this paper
• Transforming the untransformable with knockout minicircles

  2023cites this paper
• High-Efficiency Single-Cell Containment Microdevices Based on Fluid Control

  2023cites this paper
• Establishing genetic manipulation for novel strains of human gut bacteria

  2023cites this paper
• The RNA-bound proteome of MRSA reveals post-transcriptional roles for helix-turn-helix DNA-binding and Rossmann-fold proteins

  2022cites this paper
• Involvement of the Iron-Regulated Loci hts and fhuC in Biofilm Formation and Survival of Staphylococcus epidermidis within the Host

  2022cites this paper
• The host phylogeny determines viral infectivity and replication across Staphylococcus host species

  2022cites this paper
• Genomic characterization of two community-acquired methicillin-resistant Staphylococcus aureus with novel sequence types in Kenya

  2022cites this paper
• Barriers to genetic manipulation of Enterococci: Current Approaches and Future Directions

  2022cites this paper
• Exploiting transposons in the study of Staphylococcus aureus pathogenesis and virulence

  2022cites this paper
• Foodborne Origin and Local and Global Spread of Staphylococcus saprophyticus Causing Human Urinary Tract Infections

  2021cites this paper
• Methicillin-resistant Staphylococcus pseudintermedius synthesizes deoxyadenosine to cause persistent infection

  2021cites this paper
• Correlation between type IIIA CRISPR–Cas system and SCCmec in Staphylococcus epidermidis

  2021cites this paper
• From cloning to mutant in 5 days: rapid allelic exchange in Staphylococcus aureus

  2021cites this paper
• Designing multi-epitope vaccine against Staphylococcus aureus by employing subtractive proteomics, reverse vaccinology and immuno-informatics approaches

  2021cites this paper
• Conjugative DNA Transfer From E. coli to Transformation-Resistant Lactobacilli

  2021cites this paper
• Molecular Characterization of Staphylococcus aureus Plasmids Associated With Strains Isolated From Various Retail Meats

  2020cites this paper
• Prophage-Mediated Disruption of Genetic Competence in Staphylococcus pseudintermedius

  2020cites this paper
• The Accessory Gene saeP of the SaeR/S Two-Component Gene Regulatory System Impacts Staphylococcus aureus Virulence During Neutrophil Interaction

  2020cites this paper
• Posttranslational modification of Elongation Factor P from Staphylococcus aureus

  2020cites this paper
• Single-nucleotide polymorphisms in a vancomycin-resistant Staphylococcus aureus strain based on whole-genome sequencing

  2020cites this paper
• Evolution of Daptomycin Resistance in Coagulase-Negative Staphylococci Involves Mutations of the Essential Two-Component Regulator WalKR

  2019cites this paper
• A Fluorescence-based Method to Study Bacterial Gene Regulation in Infected Tissues.

  2019cites this paper
• Surface Proteins of Staphylococcus aureus

  2019cites this paper
• Functional genomics reveals extensive diversity in Staphylococcus epidermidis restriction modification systems compared to Staphylococcus aureus

  2019cites this paper
• The bactericidal mechanism of action of the SK-03-92 antibiotic against Staphylococcus aureus and the eradication of SK-03-92-induced bacterial persistence

  2019cites this paper
• Inactivation of farR Causes High Rhodomyrtone Resistance and Increased Pathogenicity in Staphylococcus aureus

  2019cites this paper
• Systematic evasion of the restriction-modification barrier in bacteria

  2019cites this paper
• Mining the Methylome Reveals Extensive Diversity in Staphylococcus epidermidis Restriction Modification

  2019cites this paper
• Determinants of Phage Host Range in Staphylococcus Species

  2019cites this paper
• Determinants of Staphylococcus aureus Colonization and Infection.“Exploring the Role of Cell Wall Anchored Proteins in Adhesion and Immune Evasion”

  2018cites this paper
• SyngenicDNA: stealth-based evasion of restriction-modification barriers during bacterial genetic engineering

  2018cites this paper
• Optimized universal protocol for electroporation of both coagulase-positive and -negative Staphylococci.

  2018cites this paper
• A horizontally gene transferred copper resistance locus confers hyper‐resistance to antibacterial copper toxicity and enables survival of community acquired methicillin resistant Staphylococcus aureus USA300 in macrophages

  2018cites this paper
• Tailor-made gene silencing of Staphylococcus aureus clinical isolates by CRISPR interference

  2018influential citation
• Comparative genomic analysis of Staphylococcus lugdunensis shows a closed pan-genome and multiple barriers to horizontal gene transfer

  2018cites this paper
• A comprehensive three-dimensional assay to assess neutrophil defense against bacteria.

  2018cites this paper
• Creation of deletion and insertion clonal complex 8 Staphylococcus aureus mutants using a common cloning vector.

  2018cites this paper
• Development of tools for genetic manipulation of Staphylococcus aureus

  2017cites this paper
• Identification of a Novel Gene argJ involved in Arginine Biosynthesis Critical for Persister Formation in Staphylococcus aureus

  2017cites this paper
• CRISPR/Cas9-based efficient genome editing in Staphylococcus aureus

  2017cites this paper
• Proteogenomics in Aid of Host–Pathogen Interaction Studies: A Bacterial Perspective

  2017cites this paper
• A Novel Spiro-Heterocyclic Compound Identified by the Silkworm Infection Model Inhibits Transcription in Staphylococcus aureus

  2017cites this paper
• New Insights into the Microbiota of Moth Pests

  2017cites this paper
• Architecture of a Species: Phylogenomics of Staphylococcus aureus.

  2017cites this paper
• Virulence Attenuation through Chemical and Genetic Manipulation of the Staphylococcus aureus ClpXP Protease

  2017cites this paper
• Restriction-Modification Systems as a Barrier for Genetic Manipulation of Staphylococcus aureus.

  2016cites this paper
• Efficient plasmid transduction to Staphylococcus aureus strains insensitive to the lytic action of transducing phage.

  2016cites this paper
• Functional characterisation of Staphylococcus pseudintermedius cell wall-associated proteins

  2016influential citation
• The remarkably multifunctional fibronectin binding proteins of Staphylococcus aureus

  2016cites this paper
• The Type 2 dUTPase of Bacteriophage ϕNM1 Initiates Mobilization of Staphylococcus aureus Bovine Pathogenicity Island 1.

  2016influential citation
• How to transform a recalcitrant Paenibacillus strain: From culture medium to restriction barrier.

  2016cites this paper
• Role of peptidoglycan deacetylase in Staphylococcus aureus virulence and survival

  2016cites this paper
• Genetic engineering of untransformable coagulase-negative staphylococcal pathogens

  2016cites this paper
• Chapter 37 – Staphylococcus aureus

  2015cites this paper
• Cell Wall-Anchored Surface Proteins of Staphylococcus aureus: Many Proteins, Multiple Functions.

  2015cites this paper
• The internal bacterial diversity of stored product pests

  2015cites this paper
• Phenotypic and Genotypic Investigation of Persistent Staphylococcus aureus Bacteraemia Isolates

  2015influential citation
• Complete Bypass of Restriction Systems for Major Staphylococcus aureus Lineages

  2015cites this paper
• Premethylation of Foreign DNA Improves Integrative Transformation Efficiency in Synechocystis sp. Strain PCC 6803

  2015cites this paper
• Improving Transformation of Staphylococcus aureus Belonging to the CC1, CC5 and CC8 Clonal Complexes

  2015cites this paper
• Engineering Cyanobacteria to Convert Carbon Dioxide to Building Blocks for Renewable Plastics

  2014cites this paper
• Population genetics and the evolution of virulence in Staphylococcus aureus.

  2014cites this paper
• Adhesion, invasion and evasion: the many functions of the surface proteins of Staphylococcus aureus

  2013cites this paper
• An update on the molecular genetics toolbox for staphylococci

  2013influential citation
• Sortase A promotes virulence in experimental Staphylococcus lugdunensis endocarditis.

  2013cites this paper
• Functional genomics of Staphylococcus aureus.

  2013cites this paper
• Functional genomics of Staphylococcus

  2013cites this paper
• The Staphylococci and Staphylococcal Pathogenesis

  2012cites this paper