Bacteriophage T4 as a Nanoparticle Platform to Display and Deliver Pathogen Antigens: Construction of an Effective Anthrax Vaccine

Li, Qin; Shivachandra, Sathish Bhadravati; Rao, Venigalla B.

doi:10.1007/978-1-4939-6869-5_15

Cited by 22 publications

(26 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…WT T4 was propagated on E. coli P301 as described previously. 46 T4(C) is a mutant containing an amber mutation at amino acid 58 of gene 42 that codes for dCMP hydroxymethylase and an amber mutation at amino acid 124 of gene 56. 30 To prevent accumulation of spontaneous revertants, the T4(C) mutant was propagated on E. coli B834 for only one generation.…”

Section: Methodsmentioning

confidence: 99%

Engineering of Bacteriophage T4 Genome Using CRISPR-Cas9

et al. 2017

Self Cite

View full text Add to dashboard Cite

Bacteriophages likely constitute the largest biomass on Earth. However, very few phage genomes have been well-characterized, the tailed phage T4 genome being one of them. Even in T4, much of the genome remained uncharacterized. The classical genetic strategies are tedious, compounded by genome modifications such as cytosine hydroxylmethylation and glucosylation which makes T4 DNA resistant to most restriction endonucleases. Here, using the type-II CRISPR-Cas9 system, we report the editing of both modified (ghm-Cytosine) and unmodified (Cytosine) T4 genomes. The modified genome, however, is less susceptible to Cas9 nuclease attack when compared to the unmodified genome. The efficiency of restriction of modified phage infection varied greatly in a spacer-dependent manner, which explains some of the previous contradictory results. We developed a genome editing strategy by codelivering into E. coli a CRISPR-Cas9 plasmid and a donor plasmid containing the desired mutation(s). Single and multiple point mutations, insertions and deletions were introduced into both modified and unmodified genomes. As short as 50-bp homologous flanking arms were sufficient to generate recombinants that can be selected under the pressure of CRISPR-Cas9 nuclease. A 294-bp deletion in RNA ligase gene rnlB produced viable plaques, demonstrating the usefulness of this editing strategy to determine the essentiality of a given gene. These results provide the first demonstration of phage T4 genome editing that might be extended to other phage genomes in nature to create useful recombinants for phage therapy applications.

show abstract

Section: Methodsmentioning

confidence: 99%

Engineering of Bacteriophage T4 Genome Using CRISPR-Cas9

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“… E. coli strains DH5α was used for cloning. E. coli strain P301 was used to propagate hoc − soc − phage T4 as described previously ( 49 – 51 ). The E. coli BL21-CodonPlus (DE3)-RIPL cells (Agilent Technologies, Santa Clara, CA) were used for expression of genes encoding target proteins.…”

Section: Methodsmentioning

confidence: 99%

“…Hoc − Soc − T4 phages were purified as described previously ( 48 , 49 ). Briefly, the propagated T4 phage on E. coli P301 was collected by centrifugation for 45 min at 25,000 × g .…”

Section: Methodsmentioning

confidence: 99%

A Bacteriophage T4 Nanoparticle-Based Dual Vaccine against Anthrax and Plague

Pan

Mahalingam

Zhu

et al. 2018

mBio

Self Cite

View full text Add to dashboard Cite

Following the deadly anthrax attacks of 2001, the Centers for Disease Control and Prevention (CDC) determined that Bacillus anthracis and Yersinia pestis that cause anthrax and plague, respectively, are two Tier 1 select agents that pose the greatest threat to the national security of the United States. Both cause rapid death, in 3 to 6 days, of exposed individuals. We engineered a virus nanoparticle vaccine using bacteriophage T4 by incorporating key antigens of both B. anthracis and Y. pestis into one formulation. Two doses of this vaccine provided complete protection against both inhalational anthrax and pneumonic plague in animal models. This dual anthrax-plague vaccine is a strong candidate for stockpiling against a potential bioterror attack involving either one or both of these biothreat agents. Further, our results establish the T4 nanoparticle as a novel platform to develop multivalent vaccines against pathogens of high public health significance.

show abstract

“…Most phages have more than one structural protein that can be used to display and, therefore, are able to link the antigen and the DC-targeting molecule to the same VLP. For example, we have shown that the two nonessential capsid proteins, Hoc and Soc, of phage T4 can be used to simultaneously display two different foreign proteins (Li et al., 2007; Shivachandra et al., 2007; Tao et al., 2017a), one an antigen and another a DC-targeting molecule (e.g., a monoclonal antibody against DEC-205) (Tao et al., 2013b).…”

Section: Applications Of Phages In Infectious Diseasementioning

confidence: 99%

Genetic Engineering of Bacteriophages Against Infectious Diseases

Chen¹,

Batra²,

Dong³

et al. 2019

Front. Microbiol.

Self Cite

135

103

View full text Add to dashboard Cite

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.

show abstract

Bacteriophage T4 as a Nanoparticle Platform to Display and Deliver Pathogen Antigens: Construction of an Effective Anthrax Vaccine

Cited by 22 publications

References 23 publications

Engineering of Bacteriophage T4 Genome Using CRISPR-Cas9

Engineering of Bacteriophage T4 Genome Using CRISPR-Cas9

A Bacteriophage T4 Nanoparticle-Based Dual Vaccine against Anthrax and Plague

Genetic Engineering of Bacteriophages Against Infectious Diseases

Contact Info

Product

Resources

About