Role of CRISPR-Cas systems and anti-CRISPR proteins in bacterial antibiotic resistance

Heliyon, Год журнала: 2024, Номер 10(14), С. e34692 - e34692

Опубликована: Июль 1, 2024

The emergence and development of antibiotic resistance in bacteria is a serious threat to global public health. Antibiotic genes (ARGs) are often located on mobile genetic elements (MGEs). They can be transferred among by horizontal gene transfer (HGT), leading the spread drug-resistant strains treatment failure. CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated genes) one many strategies have developed under long-term selection pressure restrict HGT. CRISPR-Cas systems exist about half bacterial genomes play significant role limiting resistance. On other hand, bacteriophages MGEs encode wide range anti-CRISPR proteins (Acrs) counteract immunity system. Acrs could decrease system's activity against phages facilitate acquisition ARGs virulence traits for bacteria. This review aimed assess relationship between with We also highlighted technology control prevent antibacterial system target nucleic acid sequences high accuracy reliability; therefore, it has become novel editing therapy tool CRISPR-based approaches may pave way developing smart antibiotics, which eliminate multidrug-resistant (MDR) distinguish pathogenic beneficial microorganisms. Additionally, engineered gene-containing combination antibiotics used as cutting-edge approach reduce

Язык: Английский

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PathCrisp: an innovative molecular diagnostic tool for early detection of NDM-resistant infections

Scientific Reports, Год журнала: 2025, Номер 15(1)

Опубликована: Янв. 2, 2025

The rapid and early detection of infections antibiotic resistance markers is a critical challenge in healthcare. Currently, most commercial diagnostic tools for analyzing antimicrobial patterns pathogens require elaborate culture-based testing. Our study aims to develop rapid, accurate molecular system that can be used directly from culture, thereby introducing testing conjunction with culture tests reduce turnaround time guide therapy. PathCrisp assay, combination loop-mediated isothermal amplification CRISPR-based detection, maintained at single temperature, was designed tested on clinical isolates. specificity sensitivity the assay analyzed, post which compared polymerase chain reaction (PCR) method detect New Delhi metallo-beta-lactamase (NDM) gene carbapenem-resistant enterobacteriaceae samples. demonstrated ability as few 700 copies NDM 100% concordance PCR-Sanger sequencing method, more commonly used. Additionally, lack need kit-based DNA purification step, rather crude extraction via heating, enables direct use precise, specific providing results approximately 2 h, operates constant reducing complex equipment handling. In near future, we hope this further optimized point-of-care test kit, facilitating its various healthcare settings aiding clinicians choice antibiotics

Язык: Английский

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Ultrasensitive CRISPR/Cas12a-Based System for Detection of BlaOXA-1 Gene in Antibiotic-Resistant Microorganisms

Current Issues in Molecular Biology, Год журнала: 2025, Номер 47(4), С. 238 - 238

Опубликована: Март 29, 2025

The blaOXA-1 gene encodes an oxacillin-hydrolyzing beta-lactamase of extended-spectrum (ESBL)-producing microorganisms. is found in the resistomes some Enterobacteriaceae, Morganellaceae, Pasteurellaceae, Moraxellaceae, Aeromonadaceae, Pseudomonadaceae, Yersiniaceae, and Vibrionaceae. Most ESBL detection methods, including those to detect OXA-1-producing microorganisms, are time-consuming, require specialized equipment qualified personnel. Here, we report a new CRISPR(Clustered Regularly Interspaced Short Palindromic Repeats)/Cas12a-based assay coupled with polymerase chain reaction (PCR) sensitively OXA-1-bearing PCR-coupled CRISPR/Cas12a-based fluorescence includes (i) pre-amplification step (ii) nucleic acid step. based on commonly used PCR, CRISPR/Cas12a property nonspecifically hydrolyze single-stranded DNA fluorescent reporter molecules. takes 65 min, shortened only 5 min. developed can easily single (1.25) copies efficient not model matrix but also blaOXA-1-positive We hope that our has potential improve monitoring microorganisms therefore contribute mitigating deadly global threat antibiotic-resistant

Язык: Английский

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Reinvigorating AMR resilience: leveraging CRISPR–Cas technology potentials to combat the 2024 WHO bacterial priority pathogens for enhanced global health security—a systematic review

Tropical Medicine and Health, Год журнала: 2025, Номер 53(1)

Опубликована: Апрель 2, 2025

Abstract Background Antimicrobial resistance (AMR) poses a global health threat, particularly in low- and middle-income countries (LMICs). Clustered regularly interspaced short palindromic repeats (CRISPR)–Cas system technology offers promising tool to combat AMR by targeting disabling genes WHO bacterial priority pathogens. Thus, we systematically reviewed the potential of CRISPR–Cas address AMR. Methods This systematic review adhered Preferred Reporting Items for Systematic Reviews Meta-Analyses (PRISMA) guidelines. A comprehensive literature search was conducted using Scopus PubMed databases, focusing on publications from 2014 June 2024. Keywords included “CRISPR/Cas,” “antimicrobial resistance,” “pathogen.” The eligibility criteria required original studies involving CRISPR/Cas systems that targeted Data were extracted eligible studies, qualitatively synthesized, assessed bias Joanna Briggs Institute (JBI)-standardized tool. Results 48 revealed diverse systems, including CRISPR–Cas9, CRISPR–Cas12a, CRISPR–Cas3, various genes, such as blaOXA-232, blaNDM, blaCTX-M, ermB, vanA, mecA , fosA3 blaKPC mcr-1, which are responsible carbapenem, cephalosporin, methicillin, macrolide, vancomycin, colistin, fosfomycin resistance. Some have explored role CRISPR virulence gene suppression, enterotoxin tsst1 iutA Staphylococcus aureus Klebsiella pneumoniae . Delivery mechanisms include bacteriophages, nanoparticles, electro-transformation, conjugative plasmids, demonstrate high efficiency vitro vivo. CRISPR-based diagnostic applications demonstrated sensitivity specificity, with detection limits low 2.7 × 10 2 CFU/mL, significantly outperforming conventional methods. Experimental reported significant reductions resistant populations complete suppression strains. Engineered phagemid particles plasmid-curing been shown eliminate IncF cured plasmids carrying vanA mcr-1 blaNDM 94% efficiency, restore antibiotic susceptibility. Gene re-sensitization strategies used susceptibility E. coli blaKPC-2-mediated carbapenem MDR bacteria. Whole-genome sequencing bioinformatics tools provided deeper insights into CRISPR-mediated defense mechanisms. Optimization enhanced gene-editing efficiencies, offering approach tackling high-priority Conclusions has across While promising, challenges optimizing vivo delivery, mitigating resistance, navigating ethical-regulatory barriers must be addressed facilitate clinical translation.

Язык: Английский

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Rapid molecular detection of Senecavirus A based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and CRISPR/Cas12a

Frontiers in Bioengineering and Biotechnology, Год журнала: 2025, Номер 13

Опубликована: Апрель 4, 2025

Introduction Senecavirus A (SVA), an emerging vesicular pathogen, is responsible for porcine idiopathic disease (PIVD). This closely associated with and acute neonatal piglet mortality, presenting a substantial threat to the global swine industry. At present, absence of effective drugs or vaccines treating makes accurate diagnosis SVA paramount importance prevention control disease. Methods In this study, we combined reverse transcription loop-mediated isothermal amplification (RT-LAMP) Clustered Regularly Interspaced Short Palindromic Repeats CRISPR-associated protein12a (CRISPR/Cas12a) using dual-labelled fluorescence quencher fluorescent biotin single-stranded DNA reporter molecule develop two rapid, reliable, portable visual assays: RT-LAMP-Cas12a-FQ RT-LAMP-Cas12a-FB. Results The methods exhibited comparable detection limits, 9.6 copies/μL achieved in 40 45 minutes, respectively. They did not cross-react non-target nucleic acids extracted from other related viruses showed high specificity RNA detection. Furthermore, demonstrated satisfactory performance detecting 69 adventitious samples, no significant differences that quantitative polymerase chain reaction (RT-qPCR). Discussion summary, RT-LAMP-Cas12a-FB developed are promising early routine surveillance resource-poor areas.

Язык: Английский

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Next-Gen Nano Biosensor Technologies to Monitor Carbapenem Resistance for Personalized Medicine

Indian Journal of Microbiology, Год журнала: 2024, Номер unknown

Опубликована: Июнь 20, 2024

Язык: Английский

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PathCrisp: An Innovative Molecular Diagnostic Tool for Early Detection of NDM-Resistant Infections

medRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Июль 10, 2024

Abstract Objective The rapid and early detection of infections antibiotic resistance markers is a critical challenge in healthcare. Currently, most commercial diagnostic tools for analyzing antimicrobial patterns pathogens require elaborate culture-based testing. Our study aims to develop rapid, accurate molecular system that can be used directly from culture, thereby introducing testing conjunction with culture tests reduce turnaround time (TAT) guide therapy. Methods PathCrisp assay, combination Loop-mediated Isothermal Amplification (LAMP) CRISPR-based detection, maintained at single temperature, was designed tested on clinical isolates. specificity sensitivity the assay analyzed, post which compared Polymerase Chain Reaction (PCR) method detect New Delhi metallo-beta-lactamase (NDM) gene carbapenem-resistant Enterobacteriaceae (CRE) samples. Results demonstrated ability as few 700 copies NDM 100% concordance PCR-Sanger sequencing method, more commonly used. Additionally, lack need kit-based DNA purification step, rather crude extraction via heating, enables direct use Conclusions precise, specific providing results approximately 2 hours, operates constant reducing complex equipment handling. In near future, we hope this further optimized point-of-care test kit, facilitating its various healthcare settings aiding clinicians choice antibiotics Plain language summary Resistance Carbapenem, last-line treatment, global threat. Timely diagnosis clinician decide upon treatment. However, present available methods are either expensive or have longer time. Here, study, aim tackle both limitations by developing instrument-light, called . isothermal amplification (a temperature) CRISPR/Cas diagnosis. This provides within hours temperature. validated using Carbapenem-resistant samples gene, PCR technique previously Furthermore, target when serial dilution, works (does not pure isolated DNA), NDM-positive culture.

Язык: Английский

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PathCrisp: An Innovative Molecular Diagnostic Tool for Early Detection of NDM-Resistant Infections

Research Square (Research Square), Год журнала: 2024, Номер unknown

Опубликована: Авг. 6, 2024

Objective: The rapid and early detection of infections antibiotic resistance markers is a critical challenge in healthcare. Currently, most commercial diagnostic tools for analyzing antimicrobial patterns pathogens require elaborate culture-based testing. Our study aims to develop rapid, accurate molecular system that can be used directly from culture, thereby introducing testing conjunction with culture tests reduce turnaround time (TAT) guide therapy. Methods: PathCrisp assay, combination Loop-mediated Isothermal Amplification (LAMP) CRISPR-based detection, maintained at single temperature, was designed tested on clinical isolates. specificity sensitivity the assay analyzed, post which compared Polymerase Chain Reaction (PCR) method detect New Delhi metallo-beta-lactamase (NDM) gene carbapenem-resistant Enterobacteriaceae (CRE) samples. Results: PathCrispassay demonstrated ability as few 700 copies NDM 100% concordance PCR-Sanger sequencing method, more commonly used. Additionally, lack need kit-based DNA purification step, rather crude extraction via heating, enables direct use Conclusions: PathCrisp precise, specific providing results approximately 2 hours, operates constant reducing complex equipment handling. In near future, we hope this further optimized point-of-care test kit, facilitating its various healthcare settings aiding clinicians choice antibiotics

Язык: Английский

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Critical perspectives on advancing antibiotic resistant gene (ARG) detection technologies in aquatic ecosystems

The Science of The Total Environment, Год журнала: 2024, Номер 957, С. 177775 - 177775

Опубликована: Ноя. 30, 2024

Язык: Английский

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Mitigating Antibiotic Resistance: The Utilization of CRISPR Technology in Detection

Biosensors, Год журнала: 2024, Номер 14(12), С. 633 - 633

Опубликована: Дек. 20, 2024

Antibiotics, celebrated as some of the most significant pharmaceutical breakthroughs in medical history, are capable eliminating or inhibiting bacterial growth, offering a primary defense against wide array infections. However, rise antimicrobial resistance (AMR), driven by widespread use antibiotics, has evolved into and ominous threat to global public health. Thus, creation efficient methods for detecting genes antibiotics is imperative ensuring food safety safeguarding human The clustered regularly interspaced short palindromic repeats (CRISPR) CRISPR-associated proteins (Cas) systems, initially recognized an adaptive immune mechanism bacteria archaea, have unveiled their profound potential sensor detection, transcending notable gene-editing applications. CRISPR/Cas technology employs Cas enzymes guides RNA selectively target cleave specific DNA sequences. This review offers extensive examination highlighting unique attributes applications antibiotic detection. It outlines current utilization progress toolkit identifying both nucleic acid (resistance genes) non-nucleic (antibiotic micromolecules) targets within field In addition, it examines challenges, such sensitivity specificity, future opportunities, including development point-of-care diagnostics, providing strategic insights facilitate curbing oversight antibiotic-resistance proliferation.

Язык: Английский

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