Browsing by Author "Gambill, Lauren"

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    Optogenetic control of gut bacterial metabolism to promote longevity
    (eLife Sciences Publications, Ltd, 2020) Hartsough, Lucas A.; Park, Mooncheol; Kotlajich, Matthew V.; Lazar, John Tyler; Han, Bing; Lin, Chih-Chun J.; Musteata, Elena; Gambill, Lauren; Wang, Meng C.; Tabor, Jeffrey J.; Bioengineering
    Gut microbial metabolism is associated with host longevity. However, because it requires direct manipulation of microbial metabolism in situ, establishing a causal link between these two processes remains challenging. We demonstrate an optogenetic method to control gene expression and metabolite production from bacteria residing in the host gut. We genetically engineer an Escherichia coli strain that secretes colanic acid (CA) under the quantitative control of light. Using this optogenetically-controlled strain to induce CA production directly in the Caenorhabditis elegans gut, we reveal the local effect of CA in protecting intestinal mitochondria from stress-induced hyper-fragmentation. We also demonstrate that the lifespan-extending effect of this strain is positively correlated with the intensity of green light, indicating a dose-dependent CA benefit on the host. Thus, optogenetics can be used to achieve quantitative and temporal control of gut bacterial metabolism in order to reveal its local and systemic effects on host health and aging.
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    A split ribozyme that links detection of a native RNA to orthogonal protein outputs
    (Springer Nature, 2023) Gambill, Lauren; Staubus, August; Mo, Kim Wai; Ameruoso, Andrea; Chappell, James; Bioengineering; Biosciences
    Individual RNA remains a challenging signal to synthetically transduce into different types of cellular information. Here, we describe Ribozyme-ENabled Detection of RNA (RENDR), a plug-and-play strategy that uses cellular transcripts to template the assembly of split ribozymes, triggering splicing reactions that generate orthogonal protein outputs. To identify split ribozymes that require templating for splicing, we use laboratory evolution to evaluate the activities of different split variants of the Tetrahymena thermophila ribozyme. The best design delivers a 93-fold dynamic range of splicing with RENDR controlling fluorescent protein production in response to an RNA input. We further resolve a thermodynamic model to guide RENDR design, show how input signals can be transduced into diverse outputs, demonstrate portability across different bacteria, and use RENDR to detect antibiotic-resistant bacteria. This work shows how transcriptional signals can be monitored in situ and converted into different types of biochemical information using RNA synthetic biology.