Cell-free Protein Synthesis
Cell-free Protein Synthesis - Related Videos
04:46
Synthesis of Nanodisc-Stabilized Membrane Protein Antigens Using a Cell-Free System
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2025
Source: Gilmore, S. F., et al. Cell-Free Scaled Production and Adjuvant Addition to a Recombinant Major Outer Membrane Protein from Chlamydia muridarum for Vaccine Development. J. Vis. Exp. (2022)This video demonstrates the cell-free synthesis of nanodisc-stabilized membrane protein antigens using a dual-compartment system. The device maintains optimal conditions for efficient protein synthesis, proper folding, and stable integration into nanodiscs.
07:43
Cell-Free Protein Synthesis System for Building Synthetic Cells
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Cited by 3 •
2024
This protocol describes the Cell-Free Protein Synthesis (CFPS) system used in constructing synthetic cells. It outlines key stages with representative results in different micro-compartments. The protocol aims to establish best practices for diverse laboratories in the synthetic cell community, advancing progress in synthetic cell development.
09:43
Measurement of Heme Synthesis Levels in Mammalian Cells
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Cited by 12 •
2015
Altered intracellular heme levels are associated with common diseases such as cancer. Thus, there is a need to measure heme biosynthesis levels in diverse cells. The goal of this protocol is to provide a fast and sensitive method to measure and compare the levels of heme synthesis in different cells.
15:06
Synthesis of an Intein-mediated Artificial Protein Hydrogel
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Cited by 2 •
2014
We present the synthesis of a split-intein-mediated protein hydrogel. The building blocks of this hydrogel are two protein copolymers each containing a subunit of a trimeric protein that serves as a crosslinker and one half of a split intein. Mixing of the two protein copolymers triggers an intein trans-splicing reaction, yielding a polypeptide unit that self-assembles into a hydrogel. This hydrogel is highly pH- and temperature-stable, compatible with organic solvents, and easily incorporates...
09:14
Synthesis of Protein Bioconjugates via Cysteine-maleimide Chemistry
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Cited by 8 •
2016
This protocol details the important steps required for the bioconjugation of a cysteine containing protein to a maleimide, including reagent purification, reaction conditions, bioconjugate purification and bioconjugate characterization.
FAQs
Each component supplies a distinct function: the DNA or RNA template carries the design, ribosomes perform translation, enzymes support the reaction’s molecular steps, and amino acids provide the building blocks for the protein. Energy sources and cofactors sustain these activities. Because these ingredients are supplied in an open reaction, researchers can examine and adjust the production system directly.
An open reaction gives researchers direct access to the molecular machinery rather than requiring changes to a maintained living cell. That accessibility makes genetic designs faster to prototype and allows reaction conditions to be optimized by working with the supplied template, ribosomes, enzymes, amino acids, energy sources, and cofactors. This is central to bioengineering workflows.
Production outcomes can be influenced by how the reaction is assembled and optimized, including the selected DNA or RNA template and the availability of ribosomes, enzymes, amino acids, energy sources, and cofactors. Since the system is programmable and open, researchers can vary these supplied elements or conditions to identify combinations that improve protein production for a particular design.
A practical workflow starts with a DNA or RNA template and a chosen source of molecular machinery, either a cell extract or purified components. The researcher then combines the template with ribosomes, enzymes, amino acids, energy sources, and cofactors in a reaction. This setup allows protein production to be tested without maintaining viable cells.
In bioengineering, this approach is valuable when researchers need to prototype genetic designs rapidly or produce a protein that may harm a host cell. Because viable cells do not have to be maintained, the platform separates protein production from the constraints of supporting a living host. It therefore helps evaluate designs that might be difficult to test in cells.
Cell-free protein synthesis extends beyond making a protein for direct study. Its open, programmable format supports investigations of gene expression and the construction of biosensors, diagnostics, and engineered biochemical systems. In these applications, researchers use the controllable reaction as a bioengineering platform for linking a genetic design to a measurable or engineered function.