ArticleRNA (New York, N.Y.)2023
A highly efficient human cell-free translation system.
Article in RNA (New York, N.Y.), 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 26 citations in OpenAlex.
- Eukaryotic cell-free protein synthesis: Chassis diversification, system engineering, and emerging applications.Synthetic and systems biotechnology · 2026Review
- Article
- 40S ribosomal subunits move along the 5' UTR through an eIF4A-independent mechanism in higher eukaryotes.bioRxiv : the preprint server for biology · 2026Article
- Overcoming the eIF2α Brake in Human Cell-Derived Translation Systems.bioRxiv : the preprint server for biology · 2025Article
- Synergistic Methods to Investigate Eukaryotic Translation and Its Quality Control.Journal of molecular biology · 2025Review
- A tripartite cell-free translation system to study mammalian translation.Nature protocols · 2025Review
- Half a Century of Progress: The Evolution of Wheat Germ-Based In Vitro Translation into a Versatile Protein Production Method.International journal of molecular sciences · 2025Review
- Optimizing Human Cell-Free System for Efficient Protein Production.Journal of microbiology and biotechnology · 2025Article
- BeyondChemical reviews · 2025Review
- Article
- RNA elements required for the high efficiency of West Nile virus-induced ribosomal frameshifting.Nucleic acids research · 2025Article
- Development of a highly sensitive method to detect translational infidelity.Biology methods & protocols · 2025Article
- eIF3 engages with 3'-UTR termini of highly translated mRNAs.bioRxiv : the preprint server for biology · 2024Article
- Promoting the production of challenging proteins via induced expression in CHO cells and modified cell-free lysates harboring T7 RNA polymerase and mutant eIF2α.Synthetic and systems biotechnology · 2024Article
- A highly optimized human in vitro translation system.Cell reports methods · 2024Article
- Engineering cell-free systems by chemoproteomic-assisted phenotypic screening.RSC chemical biology · 2024Article
- JUN mRNA translation regulation is mediated by multiple 5' UTR and start codon features.PloS one · 2024Article
- Article
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3 authors at 1 institution in 1 country.
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Abstract
Cell-free protein synthesis (CFPS) systems enable easy in vitro expression of proteins with many scientific, industrial, and therapeutic applications. Here we present an optimized, highly efficient human cell-free translation system that bypasses many limitations of currently used in vitro systems. This CFPS system is based on extracts from human HEK293T cells engineered to endogenously express GADD34 and K3L proteins, which suppress phosphorylation of translation initiation factor eIF2α. Overexpression of GADD34 and K3L proteins in human cells before cell lysate preparation significantly simplifies lysate preparation. We find that expression of the GADD34 and K3L accessory proteins before cell lysis maintains low levels of phosphorylation of eIF2α in the extracts. During in vitro translation reactions, eIF2α phosphorylation increases moderately in a GCN2-dependent fashion that can be inhibited by GCN2 kinase inhibitors. This new CFPS system should be useful for exploring human translation mechanisms in more physiological conditions outside the cell.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the Socratic graph.