Challenging Post-Translational Modification in the Cell-Free Protein Synthesis System DOI Open Access

Kassidy B. Porche,

Claire Lanclos,

Yong‐Chan Kwon

et al.

Published: June 21, 2023

Post-translational modifications (PTMs) represent a cornerstone in the complexity of proteome, significantly contributing to diversifying protein structure and function. PTMs can influence function, stability, localization, interactions with other molecules. Therefore, it is important when choosing expression system ensure precise incorporation during synthesis, which paramount for producing biologically active proteins. Cell-free synthesis (CFPS) has emerged as powerful platform research toolkit synthetic biology. The open nature allows reaction environment be tailored any interest promote specific PTMs, thus allowing production desired modifications. This review presents various achieved CFPS systems, providing insights into current challenges, successes, future prospects.

Language: Английский

Cell-Free Systems to Mimic and Expand Metabolism DOI Creative Commons
Blake J. Rasor, Tobias J. Erb

ACS Synthetic Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 29, 2025

Cell-free synthetic biology incorporates purified components and/or crude cell extracts to carry out metabolic and genetic programs. While protein synthesis has historically been the primary focus, more metabolism researchers are now turning toward cell-free systems either prototype pathways for cellular implementation or design new-to-nature reaction networks that incorporate environmentally relevant substrates new energy sources. The ability design, build, test enzyme combinations in vitro accelerated efforts understand bottlenecks engineer high-yielding pathways. However, only a small fraction of possibilities explored systems, from model organisms remain most common starting points. Expanding scope include organisms, alternative pathways, non-natural chemistries will enhance our bio-based chemical conversions.

Language: Английский

Citations

0
Cell-Free Synthesis System: An Accessible Platform from Biosensing to Biomanufacturing DOI
Yongbin Chen, Wenhao Xia, Fuping Lu

et al.

Microbiological Research, Journal Year: 2025, Volume and Issue: 293, P. 128079 - 128079

Published: Jan. 30, 2025

Language: Английский

Citations

0
Establishing a Cell-Free Glycoprotein Synthesis System for Enzymatic N-GlcNAcylation DOI
Madison A. DeWinter, Derek A. Wong, Regina Fernández-Morales

et al.

ACS Chemical Biology, Journal Year: 2024, Volume and Issue: 19(7), P. 1570 - 1582

Published: June 27, 2024

-linked glycosylation plays a key role in the efficacy of many therapeutic proteins. One limitation to bacterial glycoengineering human

Language: Английский

Citations

1
Carbon Negative Synthesis of Amino Acids Using a Cell-Free-Based Biocatalyst DOI Creative Commons

Shaafique Chowdhury,

Ray Westenberg,

Kimberly Wennerholm

et al.

ACS Synthetic Biology, Journal Year: 2024, Volume and Issue: 13(12), P. 3961 - 3975

Published: Nov. 21, 2024

Biological systems can directly upgrade carbon dioxide (CO

Language: Английский

Citations

1
A Cell-Free Protein Synthesis Platform to Produce a Clinically Relevant Allergen Panel DOI
Ariel Helms Thames, Clayton H. Rische, Yun Cao

et al.

ACS Synthetic Biology, Journal Year: 2023, Volume and Issue: 12(8), P. 2252 - 2261

Published: Aug. 8, 2023

Allergens are used in the clinical diagnosis (e.g., skin tests) and treatment immunotherapy) of allergic diseases. With growing interest molecular allergy diagnostics precision therapies, new tools needed for producing allergen-based reagents. As a step to address this need, we demonstrate cell-free protein synthesis approach allergen production clinically relevant panel composed common allergens spanning wide range phylogenetic kingdoms. We show that produced with can be recognized by allergen-specific immunoglobulin E (IgE), either monoclonals or patient sera. also expressed activate human cells such as peripheral blood basophils CD34+ progenitor-derived mast an IgE-dependent manner. anticipate platform will enable diagnostic therapeutic technologies, providing useful treatments both allergist patient.

Language: Английский

Citations

2
A cell-free workflow for detecting and characterizing RiPP recognition element-precursor peptide interactions DOI Creative Commons
Derek A. Wong, Zachary M. Shaver, Maria D. Cabezas

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 26, 2024

Abstract Post-translational modifications (PTMs) are important for the stability and function of many therapeutic proteins peptides. Current methods studying engineering PTM installing often suffer from low-throughput experimental techniques. Here we describe a generalizable, in vitro workflow coupling cell-free protein synthesis (CFPS) with AlphaLISA rapid expression testing proteins. We apply our to two representative classes peptide therapeutics: ribosomally synthesized post-translationally modified peptides (RiPPs) conjugate vaccines. First, demonstrate how can be used characterize binding activity RiPP recognition elements, an first step biosynthesis, integrated into biodiscovery pipeline computationally predicted products. Then, adapt study engineer oligosaccharyltransferases (OSTs) involved vaccine production, enabling identification mutant OSTs sites within carrier that enable high efficiency production In total, expect will accelerate design-build-test cycles PTMs.

Language: Английский

Citations

0
Challenging Post-translational Modifications in the Cell-free Protein Synthesis System DOI Creative Commons

Kassidy B. Porche,

Claire Lanclos,

Yong‐Chan Kwon

et al.

Synthetic biology and engineering, Journal Year: 2023, Volume and Issue: 1(1), P. 1 - 7

Published: Jan. 1, 2023

Post-translational modifications (PTMs) represent a cornerstone in the complexity of proteome, significantly contributing to diversifying protein structure and function. PTMs can considerably influence function, stability, localization, interactions with other molecules. Therefore, it is important when choosing expression system ensure precise incorporation during synthesis, which paramount for producing biologically active proteins. The cell-free synthesis (CFPS) has emerged as powerful platform research toolkit synthetic biology. open nature allows reaction environment be tailored any interest promote specific PTMs, thus allowing production desired modifications. This review presents various achieved CFPS systems, providing insights into current challenges, successes, future prospects.

Language: Английский

Citations

1
Challenging Post-Translational Modification in the Cell-Free Protein Synthesis System DOI Open Access

Kassidy B. Porche,

Claire Lanclos,

Yong‐Chan Kwon

et al.

Published: June 21, 2023

Post-translational modifications (PTMs) represent a cornerstone in the complexity of proteome, significantly contributing to diversifying protein structure and function. PTMs can influence function, stability, localization, interactions with other molecules. Therefore, it is important when choosing expression system ensure precise incorporation during synthesis, which paramount for producing biologically active proteins. Cell-free synthesis (CFPS) has emerged as powerful platform research toolkit synthetic biology. The open nature allows reaction environment be tailored any interest promote specific PTMs, thus allowing production desired modifications. This review presents various achieved CFPS systems, providing insights into current challenges, successes, future prospects.

Language: Английский

Citations

0