Engineered bacteriophages as programmable biocontrol agents

Huss, Phil; Raman, Srivatsan

doi:10.1016/j.copbio.2019.11.013

Cited by 38 publications

(25 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Despite notable exceptions, in general development of effective phage-based therapeutics is hindered by onset of bacterial resistance, resulting in low phage susceptibility. Although initial application of phages in a laboratory setting may reduce bacterial levels, the residual bacterial load remains high, causing bacteria to quickly recover after phage application ( Fister et al, 2016 ; Huss and Raman, 2020 ; Silva et al, 2014 ). A high ratio of phage to bacteria (multiplicity of infection [MOI]) may productively kill bacteria in a laboratory setting by overwhelming a host with many phages ( Abedon, 2011 ).…”

Section: Resultsmentioning

confidence: 99%

Mapping the functional landscape of the receptor binding domain of T7 bacteriophage by deep mutational scanning

Huss

Meger

Leander

et al. 2021

eLife

Self Cite

View full text Add to dashboard Cite

The interaction between a bacteriophage and its host is mediated by the phage's receptor binding protein (RBP). Despite its fundamental role in governing phage activity and host range, molecular rules of RBP function remain a mystery. Here, we systematically dissect the functional role of every residue in the tip domain of T7 phage RBP (1660 variants) by developing a high-throughput, locus-specific, phage engineering method. This rich dataset allowed us to cross compare functional profiles across hosts to precisely identify regions of functional importance, many which were previously unknown. Substitution patterns showed host-specific differences in position and physicochemical properties of mutations, revealing molecular adaptation to individual hosts. We discovered gain-of-function variants against resistant hosts and host-constricting variants that eliminated certain hosts. To demonstrate therapeutic utility, we engineered highly active T7 variants against urinary tract pathogen. Our approach presents a generalized framework for characterizing sequence-function relationships in many phage-bacterial systems.

show abstract

Section: Resultsmentioning

confidence: 99%

Mapping the functional landscape of the receptor binding domain of T7 bacteriophage by deep mutational scanning

Huss

Meger

Leander

et al. 2021

eLife

Self Cite

View full text Add to dashboard Cite

show abstract

“…The technique is based on the design of hybrid particles displaying various phage tail/tail fiber proteins with which potentially any desired bacterial host can be targeted. Besides the use of phage cocktails, it is likely that tailored phages (e.g., modification of natural phages) will be one of the clues towards realization of phage therapy [ 20 ]. Nevertheless, owing to the immense diversity of phages in nature, it will always be worthwhile to continue “phage culturomics” [ 21 ], in order to enlarge our picture of the underexplored world of phages [ 22 ].…”

Section: Resultsmentioning

confidence: 99%

Tackling Intrinsic Antibiotic Resistance in Serratia marcescens with a Combination of Ampicillin/Sulbactam and Phage SALSA

et al. 2020

View full text Add to dashboard Cite

During the antibiotic crisis, bacteriophages (briefly phages) are increasingly considered as potential antimicrobial pillars for the treatment of infectious diseases. Apart from acquired drug resistance, treatment options are additionally hampered by intrinsic, chromosomal-encoded resistance. For instance, the chromosomal ampC gene encoding for the AmpC-type β-lactamases is typically present in a number of nosocomial pathogens, including S. marcescens. In this study, phage SALSA (vB_SmaP-SALSA), with lytic activity against clinical isolates of S. marcescens, was isolated from effluent. Besides phage characterization, the aim of this study was to evaluate whether a synergistic effect between the antibiotic ampicillin/sulbactam (SAM) and phage can be achieved despite intrinsic drug resistance. Phage SALSA belongs to the Podoviridae family and genome-wide treeing analysis groups this phage within the phylogenetic radiation of T7-like viruses. The genome of Phage SALSA consists of 39,933 bp, which encode for 49 open reading frames. Phage SALSA was able to productively lyse 5 out of 20 clinical isolates (25%). A bacterial challenge with phage alone in liquid medium revealed that an initial strong bacterial decline was followed by bacterial re-growth, indicating the emergence of phage resistance. In contrast, the combination of SAM and phage, together at various concentrations, caused a complete bacterial eradication, confirmed by absorbance measurements and the absence of colony forming units after plating. The data show that it is principally possible to tackle the axiomatic condition of intrinsic drug resistance with a dual antimicrobial approach, which could be extended to other clinically relevant bacteria.

show abstract

“…It is important to highlight the potential of synthetic biology tools to rebuild phage genomes displaying enhanced biological properties [43]. Several genome engineering tools have been efficiently applied to phages [44].…”

Section: Strategies To Improve Phage Efficacymentioning

confidence: 99%

Targeting biofilms using phages and their enzymes

Azeredo

Garcı́a

Drulis-Kawa

2021

Current Opinion in Biotechnology

View full text Add to dashboard Cite

The complex biofilm architecture composed of extracellular polymeric structures (EPS) provides a protective shield to physiologically diverse bacterial cells immersed in its structure. The evolutionary interplay between bacteria and their viruses (phages) forced the latter ones to develop specific strategies to overcome the biofilm defensive barriers and kill sessile cells. Phages are equipped with a wide panel of enzyme-degrading EPS macromolecules which together are powerful weapons to combat biofilms. Antibiofilm performance can be achieved by combining phages or phage-borne enzymes with other antimicrobials such as antibiotics. Nevertheless, a variety of enzymes encoded in phage genomes still need to be explored. To advance in biofilm control strategies we must deepen the understanding of the biofilm biology itself, as well as discover and better exploit the unlimited antibacterial potential of phages.

show abstract

Engineered bacteriophages as programmable biocontrol agents

Cited by 38 publications

References 54 publications

Mapping the functional landscape of the receptor binding domain of T7 bacteriophage by deep mutational scanning

Mapping the functional landscape of the receptor binding domain of T7 bacteriophage by deep mutational scanning

Tackling Intrinsic Antibiotic Resistance in Serratia marcescens with a Combination of Ampicillin/Sulbactam and Phage SALSA

Targeting biofilms using phages and their enzymes

Contact Info

Product

Resources

About