Biology subjects

Mancuso, J. E.

Publications and source records attributed to Mancuso, J. E..

4 recordsLinked to original sources

bioRxiv2026

Bioplastic Production from simulated 100% in situ Mars resources

A sustained human presence on Mars requires local production of bulk materials, particularly polymers. Prior approaches to space biomanufacturing rely on human waste streams, Earth-sourced consumables, or complex chemical infrastructure, limiting their ability to scale. Here we demonstrate production of polyhydroxyalkanoate (PHA) bioplastic from simulated 100% martian resources: regolith-derived soluble nutrients, acetate electrochemically fixed from martian atmosphere, and water. We screened 16 candidate organisms for growth in a chemically defined Mars medium and identified Cupriavidus necator H16 and Pseudomonas putida KT2440 as promising chassis organisms. Adaptive laboratory evolution totaling more than 1012 cumulative cell divisions improved both species' tolerance to high concentrations of acetate and leached regolith. The top C. necator evolved isolate, referred to as sPL.001, produced more than three-fold higher PHA titer under simulated Mars conditions compared to its parent strain. These results establish a path to polymer production on Mars where consumable mass is derived entirely from local resources, decoupling bulk material production from Earth supply chains.

bioengineering↗

bioRxiv2026

Defined Mars Media (DMM), a chemically defined simulant of the soluble macro- and micro- nutrients in Mars regolith for use in biological research

Mars harsh yet workable surface conditions, such as manageable temperatures, availability of solar energy, and in situ resources like water ice, carbon dioxide, and mineral-rich regolith, make it a compelling target for supporting life beyond Earth. However, existing experiments testing chemical habitability in Mars conditions generally rely on leachates of physical regolith simulants, which vary in composition across simulant types, leaching conditions, and production batches. We introduce a defined Mars media (DMM) that accurately simulates the biologically relevant nutrients (nitrogen, phosphorus, and sulfur) and stressors (perchlorates, heavy metals) in Martian regolith when it is leached in water at neutral pH. We formulated DMM by combining direct rover and lander measurements from Mars with laboratory measurements of regolith simulant leachates. We validate DMM from a 1x to 20x concentrate, equivalent to 40 g/L to 800 g/L of leached regolith. Using DMM with acetate as a Mars atmosphere-derived carbon source, we grew eight bacteria, demonstrating that organisms can source all essential nutrients from Martian resources. We also demonstrate that microbial growth in DMM is robust to uncertainties in Martian regolith composition: sensitivity experiments can identify limiting trace element nutrients and toxins in DMM, and show that bacterial growth is maintained across at least an order of magnitude variation in their concentrations. This is the first defined Mars regolith media recipe containing both macro- and micro-nutrients, and designed specifically for biological experimentation. By shifting from variable leachate-based approaches to a defined aqueous analog, we enable controlled hypothesis testing of microbial survival, growth, and function in a Martian chemical environment. DMM will enable further research on astrobiology, biological in situ resource utilization, large-scale soil remediation, and terraforming. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=121 SRC="FIGDIR/small/719001v2_ufig1.gif" ALT="Figure 1"> View larger version (27K): org.highwire.dtl.DTLVardef@37408forg.highwire.dtl.DTLVardef@16fabfforg.highwire.dtl.DTLVardef@84fa43org.highwire.dtl.DTLVardef@4a7c5e_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

bioRxiv2026

BioBloom, a method for barcoded saturation mutagenesis of an entire bacterial genome

Saturation mutagenesis is a powerful tool for understanding and engineering the function of biological systems. and has been applied successfully to characterize the mutational landscape of individual proteins and genetic loci. However. it has not been applied at the whole-genome scale due to the challenges of both creating and quantifying a saturating set of mutations. Here we introduce BioBloom. a retron-based method for barcoded saturation mutagenesis at the scale of a whole bacterial genome. We constructed a barcoded BioBloom library with >99% projected sampling of saturating single - nucleotide polymorphism (SNP) mutations of the E. coli genome, and applied it to identify beneficial mutations under salt and antibiotic selection. Relative to other techniques like CRISPR-enabled mutagenesis or Adaptive Laboratory Evolution, BioBloom excels at identifying diverse causal SNPs quickly and at smaller working volumes. A barcoded, saturating mutation library is also a shared resource, and we are releasing the updated BioBloom-Ec.-2.0 library to the scientific community for broader adoption and application. Together. BioBloom makes barcoded saturation mutagenesis accessible at whole-genome scale, creating new opportunities for large-scale data collection and bacterial engineering.

microbiology↗

bioRxiv2023

A Pooled Cell Painting CRISPR Screening Platform Enables de novo Inference of Gene Function by Self-supervised Deep Learning

Pooled CRISPR screening has emerged as a powerful method of mapping gene functions thanks to its scalability, affordability, and robustness against well or plate-specific confounders present in array-based screening 1-6. Most pooled CRISPR screens assay for low dimensional phenotypes (e.g. fitness, fluorescent markers). Higher-dimensional assays such as perturb-seq are available but costly and only applicable to transcriptomics readouts 7-11. Recently, pooled optical screening, which combines pooled CRISPR screening and microscopy-based assays, has been demonstrated in the studies of the NFkB pathway, essential human genes, cytoskeletal organization and antiviral response 12-15. While the pooled optical screening methodology is scalable and information-rich, the applications thus far employ hypothesis-specific assays. Here, we enable hypothesis-free reverse genetic screening for generic morphological phenotypes by re-engineering the Cell Painting 16 technique to provide compatibility with pooled optical screening. We validated this technique using well-defined morphological genesets (124 genes), compared classical image analysis and self-supervised learning methods using a mechanism-of-action (MoA) library (300 genes), and performed discovery screening with a druggable genome library (1640 genes) 17. Across these three experiments we show that the combination of rich morphological data and deep learning allows gene networks to emerge without the need for target-specific biomarkers, leading to better discovery of gene functions.

genomics↗