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Biology subjects

Barajas, C.

Publications and source records attributed to Barajas, C..

4 recordsLinked to original sources

Pocket-Conditioned Generation and Down-Selection of Non-Oxime Reactivators

Acetylcholinesterase reactivation is a structure-constrained design problem in which chemical plausibility, access to central nervous system, pocket compatibility, and reactivation geometry must be considered together. We report a computational workflow for generating and down-selecting candidate non-oxime reactivators in the paraoxon-inhibited human acetylcholinesterase pocket. For generating the candidates, the first-round utilitezed pocket-conditioned DrugFlow, but due to solubility issues, the second-round candidates were output form FlexFlow, DrugFlow-OOD, DrugFlow preference alignment, and pocket-specified BoltzMol-1. A property-guided DrugDiff output provided a protein-unconditioned baseline. Although 93.9% of the DrugDiff subset passed Lipinski criteria for druglikeness, its median synthetic accessibility (SA) score signalled that it was difficult to synthesize compared against other generative approaches. A strict predicted blood-brain barrier and solubility gate retained 31.0% of Base DrugFlow, 61.0% of FlexFlow, 27.5% of DrugFlow-OOD, 3.8% of DrugFlow-PA, and 35.1% of BoltzMol-1. The second-round top-ten lists had lower SA scores and higher quantitative estimate of drug-likeness (QED) values than the DrugDiff shortlist, while DrugDiff had fewer aggregate heuristic violations. This work compares the best performing generative drug discovery models with suggested down sleection approaches for human acetylcholinesterase reactivation.

bioengineering↗

Machine Learning for Toxicity Prediction in Low-Sample Molecular Classes

Deep learning models such as Chemprop have advanced quantitative molecular property prediction, but their reliance on large training sets limits use in data-scarce domains. We propose a framework that fine-tunes a general baseline model trained on publicly available data on small, class-specific datasets. The resulting models retain the baseline's generalization ability while gaining class-specific accuracy and produce probabilistic outputs that capture uncertainty in the training data. We demonstrate the approach on three toxicity classes defined by a common core structure, target, or mode of action: (i) organophosphates, (ii) androgen receptor antagonists, and (iii) estrogen receptor beta antagonists. Each fine-tuned model outperforms classical machine-learning methods and the EPA TEST tool. The probabilistic nature of the predictions enables prioritization of compounds for experimental validation and seamless integration with data streams of varying quality, supporting iterative decision-making in chemical safety and drug discovery.

biochemistry↗

A burden-free gene overexpression system

Heterologous gene activation causes non-physiological burden on cellular resources that cells are unable to adjust to. Here, we introduce a feedforward controller that increases ribosome level upon activation of a gene of interest (GOI) to compensate for such a burden. The controller achieves this by activating a modified SpoT enzyme with sole hydrolysis activity, which lowers ppGpp level and thus de-represses ribosomes. Without the controller, activation of the GOI decreased growth rate by more than 50%. With the controller, we could activate the GOI to the same level without a growth rate decrease. A cell strain armed with the controller in co-culture enabled persistent population-level activation of a GOI, which could not be achieved by a strain devoid of the controller. The feedforward controller is a tunable, modular, and portable tool that for the first time allows dynamic gene activation without growth rate defects for bacterial synthetic biology applications.

synthetic biology↗

Spatial heterogeneity tunes the behavior of bacterial genetic circuits

Intracellular spatial heterogeneity is frequently observed in bacteria, where the chromosome occupies part of the cell’s volume and a circuit’s DNA often localizes within the cell. How this heterogeneity affects core processes and genetic circuits is still poorly understood. In fact, commonly used ordinary differential equation (ODE) models of genetic circuits assume a well-mixed ensemble of molecules and, as such, do not capture spatial aspects. Reaction-diffusion partial differential equation (PDE) models have been only occasionally used since they are difficult to integrate and do not provide mechanistic understanding of the effects of spatial heterogeneity. In this paper, we derive a reduced ODE model that captures spatial effects, yet has the same dimension as commonly used well-mixed models. In particular, the only difference with respect to a well-mixed ODE model is that the association rate constant of binding reactions is multiplied by a coefficient, which we refer to as the binding correction factor (BCF). The BCF depends on the size of interacting molecules and on their location when fixed in space and it is equal to unity in a well-mixed ODE model. The BCF can be used to investigate how spatial heterogeneity affects the behavior of core processes and genetic circuits. Specifically, our reduced model indicates that transcription and its regulation are more effective for genes located at the cell poles than for genes located on the chromosome. The extent of these effects depends on the value of the BCF, which we found to be close to unity. For translation, the value of the BCF is always greater than unity, it increases with mRNA size, and, with biologically relevant parameters, is substantially larger than unity. Our model has broad validity, has the same dimension as a well-mixed model, yet it incorporates spatial heterogeneity. This simple-to-use model can be used to both analyze and design genetic circuits while accounting for spatial intracellular effects.HighlightsIntracellular spatial heterogeneity modulates the effective association rate constant of binding reactions through a binding correction factor (BCF) that fully captures spatial effectsThe BCF depends on molecules size and location (if fixed) and can be determined experimentallySpatial heterogeneity may be detrimental or exploited for genetic circuit designTraditional well-mixed models can be appropriate despite spatial heterogeneityStatement of significance A general and simple modeling framework to determine how spatial heterogeneity modulates the dynamics of gene networks is currently lacking. To this end, this work provides a simple-to-use ordinary differential equation (ODE) model that can be used to both analyze and design genetic circuits while accounting for spatial intracellular effects. We apply our model to several core biological processes and determine that transcription and its regulation are more effective for genes located at the cell poles than for genes located on the chromosome and this difference increases with regulator size. For translation, we predict the effective binding between ribosomes and mRNA is higher than that predicted by a well-mixed model, and it increases with mRNA size. We provide examples where spatial effects are significant and should be considered but also where a traditional well-mixed model suffices despite severe spatial heterogeneity. Finally, we illustrate how the operation of well-known genetic circuits is impacted by spatial effects.Competing Interest StatementThe authors have declared no competing interest.View Full Text

synthetic biology↗