Phage-assisted Evolution
Phage-assisted Evolution - Related Videos
05:08
A Practical Guide to Phage- and Robotics-Assisted Near-Continuous Evolution
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2024
Phage- and Robotics-assisted Near-continuous Evolution (PRANCE) is a technique for rapid, robust protein evolution. Robotics allows the parallelization of experiments, real-time monitoring, and feedback control.
10:52
Precise Phage Mutagenesis with NgTET-Assisted CRISPR-Cas Systems
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2025
Here, we present a protocol to reduce DNA modifications in bacteriophages using the NgTET enzyme, enabling efficient and scarless CRISPR-Cas mutagenesis. This method facilitates the genetic engineering of phages for applications in biotechnology and phage therapy.
04:30
Phage Purification from Borrelia burgdorferi
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2026
Source: Eggers, C. H. Phage-Mediated Genetic Manipulation of the Lyme Disease Spirochete Borrelia burgdorferi. J. Vis. Exp. (2022).This video demonstrates the isolation of borrelial phage particles from a Borrelia burgdorferi suspension using polyethylene glycol-mediated phase separation. The sample is centrifuged, treated with a salt solution, and a molecular crowding agent to precipitate phages. Chloroform extraction is done to remove debris and concentrate phage particles. The purified phage...
03:04
Atomic Force Microscopy to Visualize Phage–Antigen Interactions
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2026
Source: Williams, S. M. et al. Novel Atomic Force Microscopy Based Biopanning for Isolation of Morphology Specific Reagents against TDP-43 Variants in Amyotrophic Lateral Sclerosis. J. Vis. Exp. (2015)This video demonstrates tapping-mode AFM to visualize phage–antigen interactions on a mica surface. It outlines the steps for antigen immobilization, phage binding, and high-resolution nanoscale imaging.
02:46
A Phage Therapy against Pseudomonas aeruginosa Infection in Zebrafish Embryos
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2025
The video demonstrates the phage therapy technique targeting Pseudomonas aeruginosa infection in cystic fibrosis zebrafish embryos. Phage therapy effectively diminishes bacterial load and mitigates proinflammatory immune responses in CF zebrafish embryos infected with bacteria, leading to reduced CF embryo lethality.
FAQs
The selection circuit makes successful phage replication dependent on activation of a required host gene, such as the infectivity factor gene III. A variant that performs the target activity more effectively triggers the circuit more reliably, producing more progeny. This couples biochemical performance to reproductive success, allowing functional differences among variants to become population-level enrichment over successive generations.
Persistence depends on how effectively a variant activates the host selection circuit relative to competing variants. Improved activity supports more successful phage reproduction, while weaker variants reproduce less effectively and are progressively removed from the flowing culture. The balance between replication and washout therefore translates functional performance into changes in variant abundance.
Continuous flow maintains an ongoing competition among phage variants while removing organisms that do not reproduce successfully enough to remain in the system. Because selection proceeds across many generations with limited manual intervention, beneficial activity can accumulate rapidly rather than requiring repeated cycles of recovery, testing, and reseeding. This makes the approach suited to accelerated directed evolution.
A workable setup coordinates phage carrying the gene or genetic system being evolved, engineered host cells, and a selection circuit that connects the target activity to a required replication function. The culture must also support continuous flow, so successful variants can reproduce while less functional variants are washed out. These components together impose the intended evolutionary pressure.
Researchers first place the gene or genetic system of interest in the phage population, then design the host-cell selection circuit so that the desired activity activates a gene required for phage replication. Variants are maintained under continuous-flow selection, where more functional forms gain a reproductive advantage. The resulting population can therefore become enriched for improved target activity.
The method has been applied to several classes of molecular tools, including enzymes, polymerases, and genome-editing components. Its value is not limited to increasing a single measured activity; selection can support the development of tools with new or improved functions when those functions can be connected to the phage replication circuit. This broadens its relevance across bioengineering research.