Phage Genetic Engineering Using CRISPR–Cas Systems

Hatoum-Aslan, Asma

doi:10.3390/v10060335

Cited by 64 publications

(43 citation statements)

References 75 publications

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“…Previous studies have used phages in the preparations of genetically engineered cocktails according to receptor-binding proteins, generally TFP, with a wide range of recognition capabilities [8,43]. Many people also make use of gene-editing technology to artificially modify TFPs to detect and treat pathogenic bacteria [44][45][46][47]. Tail fiber proteins on the tail fibers of bacteriophages can be massively generated through genetic engineering and can be used as suitable probes to target their host bacteria.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of AbTJ, an Acinetobacter baumannii Phage, and Functional Identification of Its Receptor-Binding Modules

Kang

et al. 2020

Viruses

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A. baumannii is an opportunistic pathogen and a major cause of various community-acquired infections. Strains of this species can be resistant to multiple antimicrobial agents, leaving limited therapeutic options, also lacking in methods for accurate and prompt diagnosis. In this context, AbTJ, a novel phage that infects A. baumannii MDR-TJ, was isolated and characterized, together with its two tail fiber proteins. Morphological analysis revealed that it belongs to Podoviridae family. Its host range, growth characteristics, stability under various conditions, and genomic sequence, were systematically investigated. Bioinformatic analysis showed that AbTJ consists of a circular, double-stranded 42670-bp DNA molecule which contains 62 putative open reading frames (ORFs). Genome comparison revealed that the phage AbTJ is related to the Acinetobacter phage Ab105-1phi (No. KT588074). Tail fiber protein (TFPs) gp52 and gp53 were then identified and confirmed as species-specific proteins. By using a combination of bioluminescent methods and magnetic beads, these TFPs exhibit excellent specificity to detect A. baumannii. The findings of this study can be used to help control opportunistic infections and to provide pathogen-binding modules for further construction of engineered bacteria of diagnosis and treatment.

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Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of AbTJ, an Acinetobacter baumannii Phage, and Functional Identification of Its Receptor-Binding Modules

Kang

et al. 2020

Viruses

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“…In general, natural and engineered phage cocktails dominate our sample. New genetic tools such as CRISPR-Cas systems are used to genetically engineer phages that infect diverse hosts 46 . Phages are also used as species-specific carriers for a variety of potential antibacterial payloads 47 or CRISPR-Cas-based RNA-guided nucleases targeted at resistance or virulence determinants 48,49 .…”

Section: Phage and Phage-derived Peptidesmentioning

confidence: 99%

The global preclinical antibacterial pipeline

Theuretzbacher¹,

et al. 2019

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| Antibacterial resistance is a great concern and requires global action. A critical question is whether enough new antibacterial drugs are being discovered and developed. A review of the clinical antibacterial drug pipeline was recently published, but comprehensive information about the global preclinical pipeline is unavailable. This Review focuses on discovery and preclinical development projects and has found, as of 1 May 2019, 407 antibacterial projects from 314 institutions. The focus is on Gram-negative pathogens, particularly bacteria on the WHO priority bacteria list. The preclinical pipeline is characterized by high levels of diversity and interesting scientific concepts, with 135 projects on direct-acting small molecules that represent new classes, new targets or new mechanisms of action. There is also a strong trend towards non-traditional approaches, including diverse antivirulence approaches, microbiome-modifying strategies, and engineered phages and probiotics. The high number of pathogen-specific and adjunctive approaches is unprecedented in antibiotic history. Translational hurdles are not adequately addressed yet, especially development pathways to show clinical impact of nontraditional approaches. The innovative potential of the preclinical pipeline compared with the clinical pipeline is encouraging but fragile. Much more work , focus and funding are needed for the novel approaches to result in effective antibacterial therapies to sustainably combat antibacterial resistance.

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“…Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas system, firstly discovered at the end of last century, is an immune system of prokaryote to counter the invasions. Recently, it was adapted for genome engineering in many organisms including phages (Figure 2; Box et al., 2016; Bari et al., 2017; Tao et al., 2017b; Hatoum-Aslan, 2018; Hupfeld et al., 2018; Knott and Doudna, 2018; Schilling et al., 2018; Shen et al., 2018). The effector complexes of CRISPR-Cas system contain two main components, Cas proteins and CRISPR RNA (crRNA).…”

Section: Phage Genome Engineeringmentioning

confidence: 99%

Genetic Engineering of Bacteriophages Against Infectious Diseases

Chen¹,

Batra²,

Dong³

et al. 2019

Front. Microbiol.

135

103

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Bacteriophages (phages) are the most abundant and widely distributed organisms on Earth, constituting a virtually unlimited resource to explore the development of biomedical therapies. The therapeutic use of phages to treat bacterial infections (“phage therapy”) was conceived by Felix d’Herelle nearly a century ago. However, its power has been realized only recently, largely due to the emergence of multi-antibiotic resistant bacterial pathogens. Progress in technologies, such as high-throughput sequencing, genome editing, and synthetic biology, further opened doors to explore this vast treasure trove. Here, we review some of the emerging themes on the use of phages against infectious diseases. In addition to phage therapy, phages have also been developed as vaccine platforms to deliver antigens as part of virus-like nanoparticles that can stimulate immune responses and prevent pathogen infections. Phage engineering promises to generate phage variants with unique properties for prophylactic and therapeutic applications. These approaches have created momentum to accelerate basic as well as translational phage research and potential development of therapeutics in the near future.

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Phage Genetic Engineering Using CRISPR–Cas Systems

Cited by 64 publications

References 75 publications

Isolation and Characterization of AbTJ, an Acinetobacter baumannii Phage, and Functional Identification of Its Receptor-Binding Modules

Isolation and Characterization of AbTJ, an Acinetobacter baumannii Phage, and Functional Identification of Its Receptor-Binding Modules

The global preclinical antibacterial pipeline

Genetic Engineering of Bacteriophages Against Infectious Diseases

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