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Self-cleaving ribozymes are important tools in synthetic biology, biomanufacturing, and nucleic acid therapeutics. These broad applications deploy ribozymes in many genetic and environmental contexts, which can influence activity. Thus, accurate
d'Oelsnitz Simon, Wantae Kim, Haley Hardtke, Svetlana Ikonomova, Nina Alperovich, Olga Vasilyeva, Michael James, Eric Zigon, Charlie Johnson, Andrew Ellington, Quincey Justman, Michael Springer, Jessie Zhang, Pamela Silver, David Ross
Biocatalysts are championed for their exquisite stereochemistry, but slow chromatographic separations necessary to measure enantiomeric excess can bottleneck their development. Prokaryotic transcription factors can address this limitation by transducing
Molecular biosensors that accurately measure protein concentrations without external equipment are critical for solving numerous problems in diagnostics and therapeutics. Modularly transducing the binding of protein antibodies, protein switches or aptamers
Takeshi Matsui, Po-Hsiang Hung, Han Mei, Xianan Liu, Fangfei Li, John Collins, Weiyi Li, Darach Miller, Neil Wilson, Esteban Toro, Geoffrey Taghon, Gavin Sherlock, Sasha Levy
To reduce the operational friction and scale DNA engineering, we report here an in vivo DNA assembly technology platform called SCRIVENER (Sequential Conjugation and Recombination for In Vivo Elongation of Nucleotides with low ERrors). SCRIVENER combines
G protein-coupled receptors (GPCRs) are the largest family of transmembrane receptors in humans. Over 800 GPCRs regulate the (patho)biology of every organ, tissue, and cell type. Consequently, GPCRs are the most prominent therapeutic targets in medicine
Carl Simon Jr., Anne L. Plant, Catherine Zander, Erich Bozenhardt, Christina Celluzzi, David Dobnik, Melanie Grant, John Hughes, Uma Lakshmipathy, Kok-Seong Lim, Laura Montgomery, Hesham Nawar, Thiana Nebel, Linda Peltier, James Sherley, Rouzbeh Taghizadeh, Eddie Tan, Sandrine Vessillier
One of the most challenging aspects of cell and gene therapy products (CGTPs) is defining mechanism of action (MOA), potency and efficacy of the product. This perspective examines these concepts and presents helpful ways to think about them through the
Prokaryotic transcription factors can be repurposed into biosensors for the ligand-inducible control of gene expression, but the landscape of chemical ligands for which biosensors exist is extremely limited. To expand this landscape, we developed Ligify, a
Samuel Schaffter, Molly Wintenberg, Terence Murphy, Elizabeth Strychalski
Cotranscriptionally encoded RNA strand displacement (ctRSD) circuits are an emerging tool for programmable molecular computation, with potential applications spanning in vitro diagnostics to continuous computation inside living cells. In ctRSD circuits
Eun Jin Lee, Paul Salipante, Anthony Kotula, Dana Graves, Stella Alimperti, Yoontae Kim
Epithelial cell function is modulated by mechanical forces imparted by the extracellular environment. The transmission of forces onto the cytoskeleton by modalities such as mechanical stress and matrix stiffness is necessary to address by the development
Genome editing is a rapidly evolving biotechnology with the potential to transform many sectors of industry such as agriculture, biomanufacturing, and medicine. This technology is enabled by an ever-growing portfolio of biomolecular reagents that span the
Eugenia Romantseva, Nina Alperovich, David Ross, Steven Lund, Elizabeth Strychalski
DNA templates for protein expression remain an unexplored source of variability in the performance of cell-free expression (CFE) systems for protein production. To characterize this variability, we investigated the effects of two common DNA extraction
Microbial cell culture is one of the most commonly performed protocols for synthetic biology, and laboratories are increasingly using 96-well plates and laboratory automation systems for cell culture. Here we describe a method for reproducible microbial
Large-scale measurements linking genetic background to biological function have drivena need for models that can incorporate these data for reliable predictions and insightinto the underlying biophysical system. Recent modeling efforts, however, prioritize
Drew S. Tack, Peter Tonner, Abe Pressman, Nathanael David Olson, Eugenia Romantseva, Nina Alperovich, Olga Vasilyeva, David J. Ross, Sasha F. Levy
Allostery is a fundamental biophysical mechanism where the activity of a biomolecule is regulated by the binding of a ligand. Despite playing a central role in many biological processes, a quantitative understanding of allostery is lacking. To
Camille Lombard-Banek, Kerstin Pohl, Edward Kwee, John T. Elliott, John E. Schiel
Mass Spectrometry (MS)-based proteomic measurements are uniquely poised to impact the development of cell and gene therapies. With the adoption of rigorous instrumental performance controls (PQs), large-scale proteomics can move from a research to a
Engineered molecular circuits that process information in biological systems could address emerging human health and biomanufacturing needs. However, such circuits can be difficult to rationally design and scale. DNA-based strand displacement reactions
A protocol for increasing soluble protein expression by fusing the chaperone GroEL apical domain with a gene of interest is described herein. GroEL apical domain, the minichaperone that functions independently of GroES and ATP in protein folding, is cloned
The move from reading to writing the human genome offers new opportunities to improve human health. The United States National Institutes of Health (NIH) Somatic Cell Genome Editing (SCGE) Consortium aims to accelerate the development of safer and more