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Biology subjects

Michael Baym

Publications and source records attributed to Michael Baym.

3 recordsLinked to original sources

Long-term monitoring of inflammation in the mammalian gut using programmable commensal bacteria

Inflammation in the gut, caused by infection and autoimmunity, remains challenging to effectively detect, monitor, and treat. Here, we engineer a commensal mouse E. coli strain to record exposure to tetrathionate, a downstream product of reactive oxygen species generated during inflammation. Using these programmed bacteria to sense in situ levels we show that tetrathionate accompanies inflammation during Salmonella-induced colitis in mice and is elevated in an inflammatory bowel disease mouse model. We demonstrate long-term genetic stability and associated robust function of synthetic genetic circuits in bacteria colonizing the mammalian gut. These results demonstrate the potential for engineered bacteria to stably and reliably probe pathophysiological processes for which traditional diagnostics may not be feasible or cost-effective.\n\nOne sentence summaryEngineered bacteria record an inflammatory response in an IBD mouse model and are genetically stable during long-term growth in the mouse gut.

Synthetic Biology

Rapid Construction of a Whole-genome Transposon Insertion Collection for Shewanella oneidensis by Knockout Sudoku

Whole-genome knockout collections are invaluable for connecting gene sequence to function, yet traditionally they have needed an extraordinary technical effort to construct. Knockout Sudoku is a new method for directing the construction and purification of a curated whole-genome collection of singlegene disruption mutants generated by transposon mutagenesis. Using a simple combinatorial pooling scheme, a highly oversampled collection of transposon mutants can be condensed into a next-generation sequencing library in a single day. The identities of the mutants in the collection are then solved by a predictive algorithm based on Bayesian inference, allowing for rapid curation and validation. Starting from a progenitor collection of 39,918 transposon mutants, we compiled a quality-controlled knockout collection of the electroactive microbe Shewanella oneidensis MR-1 containing representatives for 3,667 genes. High-throughput kinetic measurements on this collection provide a comprehensive view of multiple extracellular electron transfer pathways operating in parallel.

Genetics

Inexpensive Multiplexed Library Preparation for Megabase-Sized Genomes

Whole-genome sequencing has become an indispensible tool of modern biology. However, the cost of sample preparation relative to the cost of sequencing remains high, especially for small genomes where the former is dominant. Here we present a protocol for the rapid and inexpensive preparation of hundreds of multiplexed genomic libraries for Illumina sequencing. By carrying out the Nextera tagmentation reaction in small volumes, replacing costly reagents with cheaper equivalents, and omitting unnecessary steps, we achieve a cost of library preparation of $8 per sample, approximately 6 times cheaper than the widely-used Nextera XT protocol. Furthermore, our procedure takes less than 5 hours for 96 samples and uses nanograms of genomic DNA. Many hundreds of samples can then be pooled on the same HiSeq lane via custom barcodes. Our method is especially useful for re-sequencing of large numbers of full microbial or viral genomes, including those from evolution experiments, genetic screens, and environmental samples.

Genomics