ABSTRACT
Recent progress in synthetic biology has enabled the design of complex genetic circuits that interface with innate cellular functions, such as gene transcription, and control user-defined outputs. Implementing these genetic networks in mammalian cells, however, is a cumbersome process that requires several steps of optimization and benefits from the use of predictive modeling. Combining deterministic mathematical models with software-based numerical computing platforms allows researchers to quickly design, evaluate, and optimize multiple circuit topologies to establish experimental constraints that generate the desired control systems. In this chapter, we present a systematic approach based on predictive mathematical modeling to guide the design and construction of gene activity-based sensors. This approach enables user-driven circuit optimization through iterations of sensitivity analyses and parameter scans, providing a universal method to engineer sense and respond cells for diverse applications.
Subject(s)
Gene Regulatory Networks , Software , Animals , Humans , Computer Simulation , Research Personnel , Synthetic Biology , MammalsABSTRACT
The production of high-quality recombinant proteins is critical to maintaining a continuous supply of biopharmaceuticals, such as therapeutic antibodies. Engineering mammalian cell factories presents a number of limitations typically associated with the proteotoxic stress induced upon aberrant accumulation of off-pathway protein folding intermediates, which eventually culminate in the induction of apoptosis. In this review, we will discuss advances in cell engineering and their applications at different hierarchical levels of control of the expression of recombinant proteins, from transcription and translational to posttranslational modifications and subcellular trafficking. We also highlight challenges and unique opportunities to apply modern synthetic biology tools to the design of programmable cell factories for improved biomanufacturing of therapeutic proteins.
Subject(s)
Cell Engineering , Synthetic Biology , Animals , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Protein Processing, Post-Translational , Metabolic Engineering , Mammals/metabolismABSTRACT
The use of bacteriophages as antimicrobial agents represents a promising alternative for the control of pathogenic bacteria. Here, we present the complete genome sequences of two novel Salmonella enterica lytic bacteriophages, NBSal006 and NBSal007, candidates for Salmonella biocontrol.
ABSTRACT
The emergence of antibiotic resistance has raised serious concerns within scientific and medical communities, and has underlined the importance of developing new antimicrobial agents to combat such infections. Bacteriophages, naturally occurring bacterial viruses, have long been characterized as promising antibiotic alternatives. Although bacteriophages hold great promise as medical tools, clinical applications have been limited by certain characteristics of phage biology, with structural fragility under the high temperatures and acidic environments of therapeutic applications significantly limiting therapeutic effectiveness. This study presents and evaluates the efficacy of a new accelerated evolution platform, chemically accelerated viral evolution (CAVE), which provides an effective and robust method for the rapid enhancement of desired bacteriophage characteristics. Here, our initial use of this methodology demonstrates its ability to confer significant improvements in phage thermal stability. Analysis of the mutation patterns that arise through CAVE iterations elucidates the manner in which specific genetic modifications bring forth desired changes in functionality, thereby providing a roadmap for bacteriophage engineering.
Subject(s)
Anti-Bacterial Agents/pharmacology , Bacteriophages/genetics , Drug Resistance, Microbial/genetics , Genetic Engineering/methods , Animals , Evolution, Molecular , Gene Expression Regulation, Viral , Humans , Hydrogen-Ion Concentration , Phage Therapy , TemperatureABSTRACT
Salmonella enterica is a Gram-negative bacterium, recognized as one of the most important foodborne pathogens in the world. Bacteriophages represent a promising alternative to the biocontrol of Salmonella Here, we report the isolation of five Salmonella bacteriophages, the sequencing of their full genomes, and initial genomic characterization.
