Biology subjects

Riley, E. M.

Publications and source records attributed to Riley, E. M..

3 recordsLinked to original sources

bioRxiv2025

A comprehensive genetic toolkit for Zymomonas mobilis; allowing rapid, high-resolution, combinatorial engineering of metabolic pathways.

The limited availability of reliable genetic tools has constrained the precise engineering of Zymomonas mobilis, a promising bacterium for the sustainable bioproduction of fuels and commodity chemicals. Here, we present a comprehensive, modular genetic toolkit that enables high-resolution, combinatorial control of gene expression in Z. mobilis. We constructed and characterised a synthetic promoter library spanning the broadest dynamic range reported for this organism, alongside a suite of strong transcriptional terminators that effectively insulate genetic cassettes and prevent transcriptional readthrough. To achieve predictable translation across diverse genetic contexts, we further implemented bicistronic design (BCD) elements, allowing context-independent control of expression. Promoter activities were shown to be stable under both aerobic and anaerobic conditions, highlighting the robustness of the regulatory parts across physiologically relevant environments. The toolkit was integrated into the Start-Stop Assembly system, facilitating high-throughput construction of multi-gene pathways. We demonstrate its utility through the rapid assembly and testing of the possible expression space of genes encoding a 2,3-butanediol biosynthetic pathway. Collectively, this genetic toolkit now allows the high-resolution engineering of multi-enzyme pathways in Z. mobilis for industrial biotechnology applications.

synthetic biology↗

bioRxiv2022

The genetics of immune and infection phenotypes in wild mice, Mus musculus domesticus.

Wild animals are under constant threat from a wide range of micro- and macroparasites in their environment. Animals make immune responses against parasites, and these are important in affecting the dynamics of parasite populations. Individual animals vary in their anti-parasite immune responses. Genetic polymorphism of immune-related loci contributes to inter-individual differences in immune responses, but most of what we know in this regard comes from studies of humans or laboratory animals; there are very few such studies of wild animals naturally infected with parasites. Here we have investigated the effect of polymorphism in immune-related loci (the MHC, and genes coding for cytokines and Toll-like receptors) on a wide range of immune and infection phenotypes in UK wild house mice, Mus musculus domesticus. We found strong effects of polymorphisms in various MHC and cytokine coding loci on both immune measures (antibody concentration and cytokine production) and on infection phenotypes (infection with mites, worms and viruses). Our study provides a comprehensive view of how polymorphism of immune-related loci affects immune and infection phenotypes in naturally infected wild rodent populations.

ecology↗

bioRxiv2019

Natural Killer cells dampen the pathogenic features of recall responses to influenza infection.

Despite evidence of augmented Natural Killer (NK) cell responses after influenza vaccination, the role of these cells in vaccine-induced immunity remains unclear. Here, we hypothesized that NK cells might increase viral clearance but possibly at the expense of increased severity of pathology. On the contrary, we found that NK cells serve a homeostatic role during influenza virus infection of vaccinated mice, allowing viral clearance with minimal pathology. Using a diphtheria toxin receptor transgenic mouse model, we were able to specifically deplete NKp46+ NK cells through the administration of diphtheria toxin. Using this model, we assessed the effect of NK cell depletion prior to influenza challenge in vaccinated and unvaccinated mice. NK-depleted, vaccinated animals lost significantly more weight after viral challenge than vaccinated NK intact animals, indicating that NK cells ameliorate disease in vaccinated animals. However, there was also a significant reduction in viral load in NK-depleted, unvaccinated animals indicating that NK cells also constrain viral clearance. Depletion of NK cells after vaccination, but 21 days before infection, did not affect viral clearance or weight loss - indicating that it is the presence of NK cells during the infection itself that promotes homeostasis. Further work is needed to identify the mechanism(s) by which NK cells regulate adaptive immunity in influenza-vaccinated animals to allow efficient and effective virus control whilst simultaneously minimizing inflammation and pathology.

immunology↗