ACS Synthetic Biology

Journal Article

Point-of-Care Peptide Hormone Production Enabled by Cell-Free Protein Synthesis

DeWinter, Madison A.; Thames, Ariel Helms; Guerrero, Laura; Kightlinger, Weston; Karim, Ashty S.;

2023 ACS Synthetic Biology

In resource-limited settings, it can be difficult to safely deliver sensitive biologic medicines to patients due to cold chain and infrastructure constraints. Point-of-care drug manufacturing could circumvent these challenges since medicines could be produced locally and used on-demand. Toward...

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Journal Article

Indirect Enrichment of Desirable, but Less Fit Phenotypes, from a Synthetic Microbial Community Using Microdroplet Confinement

Ganiga Prabhakar, Ramya; Fan, Gaoyang; Alnahhas, Razan N.; Hirning, Andrew J.; Bennett, Matthew R.;

2023 ACS Synthetic Biology

doi:10.1021/acssynbio.3c00008

Spatial structure within microbial communities can provide nearly limitless opportunities for social interactions and are an important driver for evolution. As metabolites are often molecular signals, metabolite diffusion within microbial communities can affect the composition and dynamics of...

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Journal Article

Optimization of Hydrogenobyrinic Acid Synthesis in a Cell-Free Multienzyme Reaction by Novel S‑Adenosyl-methionine Regeneration

Xiao, Kaixing; Kang, Qian; Xiang, Mengjie; Gong, Dachun; Fang, Huan;

2023 ACS Synthetic Biology

doi:10.1021/acssynbio.3c00109

Hydrogenobyrinic acid, a modified tetrapyrrole composed of eight five-carbon compounds, is a key intermediate and central framework of vitamin B12. Synthesis of hydrogenobyrinic acid requires eight S-adenosyl-methionine working as the methyl group donor catalyzed by 12 enzymes including six...

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Journal Article

Assembly of Metabolons in Yeast Using Cas6-Mediated RNA Scaffolding

Pham, Anhuy; Bassett, Shane; Chen, Wilfred; Da Silva, Nancy A.

2023 ACS Synthetic Biology

doi:10.1021/acssynbio.2c00650 pmid:36920425

Cells often localize pathway enzymes in close proximity to reduce substrate loss via diffusion and to ensure that carbon flux is directed toward the desired product. To emulate this strategy for the biosynthesis of heterologous products in yeast, we have taken advantage of the highly specific...

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Journal Article

Directed Evolution of Aerotolerance in Sulfide-Dependent Thiazole Synthases

Gelder, Kristen Van; Oliveira-Filho, Edmar R.; García-García, Jorge Donato; Hu, You; Bruner, Steven D.;

2023 ACS Synthetic Biology

doi:10.1021/acssynbio.2c00512 pmid:36920242

Sulfide-dependent THI4 thiazole synthases could potentially be used to replace plant cysteine-dependent suicide THI4s, whose high protein turnover rates make thiamin synthesis exceptionally energy-expensive. However, sulfide-dependent THI4s are anaerobic or microoxic enzymes and hence unadapted...

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Journal Article

Standard Intein Gene Expression Ramps (SIGER) for Protein-Independent Expression Control

Fages-Lartaud, Maxime; Mueller, Yasmin; Elie, Florence; Courtade, Gaston; Hohmann-Marriott, Martin Frank

2023 ACS Synthetic Biology

doi:10.1021/acssynbio.2c00530 pmid:36920366

Coordination of multigene expression is one of the key challenges of metabolic engineering for the development of cell factories. Constraints on translation initiation and early ribosome kinetics of mRNA are imposed by features of the 5′UTR in combination with the start of the gene, referred to...

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Journal Article

High-Density Guide RNA Tiling and Machine Learning for Designing CRISPR Interference in Synechococcus sp. PCC 7002

Dallo, Tessa; Krishnakumar, Raga; Kolker, Stephanie D.; Ruffing, Anne M.

2023 ACS Synthetic Biology

doi:10.1021/acssynbio.2c00653 pmid:36893454

While CRISPRi was previously established in Synechococcus sp. PCC 7002 (hereafter 7002), the design principles for guide RNA (gRNA) effectiveness remain largely unknown. Here, 76 strains of 7002 were constructed with gRNAs targeting three reporter systems to evaluate features that impact gRNA...

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Journal Article

Marine Biofilm Engineered to Produce Current in Response to Small Molecules

Bird, Lina J.; Leary, Dagmar H.; Hervey, Judson; Compton, Jaimee; Phillips, Daniel;

2023 ACS Synthetic Biology

doi:10.1021/acssynbio.2c00417

Engineered electroactive bacteria have potential applications ranging from sensing to biosynthesis. In order to advance the use of engineered electroactive bacteria, it is important to demonstrate functional expression of electron transfer modules in chassis adapted to operationally relevant...

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Journal Article

Performance Prediction of Fundamental Transcriptional Programs

Milner, Prasaad T.; Zhang, Ziqiao; Herde, Zachary D.; Vedire, Namratha R.; Zhang, Fumin;

2023 ACS Synthetic Biology

doi:10.1021/acssynbio.2c00593 pmid:36935615

Transcriptional programming leverages systems of engineered transcription factors to impart decision-making (e.g., Boolean logic) in chassis cells. The number of components used to construct said decision-making systems is rapidly increasing, making an exhaustive experimental evaluation of...

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Journal Article

A Genetic Engineering Toolbox for the Lignocellulolytic Anaerobic Gut Fungus Neocallimastix frontalis

Hooker, Casey A.; Hanafy, Radwa; Hillman, Ethan T.; Muñoz Briones, Javier; Solomon, Kevin V.

2023 ACS Synthetic Biology

doi:10.1021/acssynbio.2c00502 pmid:36920337

Anaerobic fungi are powerful platforms for biotechnology that remain unexploited due to a lack of genetic tools. These gut fungi encode the largest number of lignocellulolytic carbohydrate active enzymes (CAZymes) in the fungal kingdom, making them attractive for applications in renewable energy...

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