Engineered Mammalian Chromosomes in Cellular Protein Production: Future Prospects

2011 ◽  
pp. 217-238 ◽  
Author(s):  
Malcolm L. Kennard
Keyword(s):  
Protein Production ◽  
Cellular Protein ◽  
Future Prospects

scholarly journals Community control in cellular protein production: consequences for amino acid starvation

2015 ◽  
Vol 373 (2056) ◽  
pp. 20150107 ◽  
Author(s):  
Frank S. Heldt ◽  
Chris A. Brackley ◽  
Celso Grebogi ◽  
Marco Thiel
Keyword(s):  
Amino Acid ◽  
Protein Production ◽  
Control Mechanism ◽  
Supply And Demand ◽  
Mrna Translation ◽  
Cellular Protein ◽  
Amino Acid Starvation ◽  
Community Control ◽  
Trna Species ◽  
A Cell

Deprivation of essential nutrients can have stark consequences for many processes in a cell. We consider amino acid starvation, which can result in bottlenecks in mRNA translation when ribosomes stall due to lack of resources, i.e. tRNAs charged with the missing amino acid. Recent experiments also show less obvious effects such as increased charging of other (non-starved) tRNA species and selective charging of isoaccepting tRNAs. We present a mechanism which accounts for these observations and shows that production of some proteins can actually increase under starvation. One might assume that such responses could only be a result of sophisticated control pathways, but here we show that these effects can occur naturally due to changes in the supply and demand for different resources, and that control can be accomplished through selective use of rare codons. We develop a model for translation which includes the dynamics of the charging and use of aminoacylated tRNAs, explicitly taking into account the effect of specific codon sequences. This constitutes a new control mechanism in gene regulation which emerges at the community level, i.e. via resources used by all ribosomes.

Biopharmaceutical protein production bySaccharomyces cerevisiae: current state and future prospects

2014 ◽  
Vol 2 (2) ◽  
pp. 167-182 ◽  
Author(s):  
Mingtao Huang ◽  
Jichen Bao ◽  
Jens Nielsen
Keyword(s):  
Protein Production ◽  
Future Prospects ◽  
Current State

scholarly journals Improving genomically recoded Escherichia coli for the production of proteins containing non-canonical amino acids

2021 ◽  
Author(s):  
Jessica G. Perez ◽  
Erik D. Carlson ◽  
Oliver Weisser ◽  
Camila Kofman ◽  
Kosuke Seki ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Protein Production ◽  
Fluorescent Protein ◽  
Cellular Protein ◽  
Release Factor ◽  
Wild Type ◽  
Genetic Encoding ◽  
Lysine Residues ◽  
Green Fluorescent

AbstractA genomically recoded Escherichia coli strain that lacks all amber codons and release factor 1 (C321.ΔA) enables efficient genetic encoding of chemically diverse, non-canonical amino acids (ncAAs) into proteins. While C321.ΔA has opened new opportunities in chemical and synthetic biology, this strain has not been optimized for protein production, limiting its utility in widespread industrial and academic applications. To address this limitation, we describe the construction of a series of genomically recoded organisms that are optimized for cellular protein production. We demonstrate that the functional deactivation of nucleases (e.g., rne, endA) and proteases (e.g., lon) increases production of wild-type superfolder green fluorescent protein (sfGFP) and sfGFP containing two ncAAs up to ∼5-fold. Additionally, we introduce a genomic IPTG-inducible T7 RNA polymerase (T7RNAP) cassette into these strains. Using an optimized platform, we demonstrated the ability to introduce 2 identical N6-(propargyloxycarbonyl)-L-Lysine residues site specifically into sfGFP with a 17-fold improvement in production relative to the parent. We envision that our library of organisms will provide the community with multiple options for increased expression of proteins with new and diverse chemistries.

scholarly journals Successful reprogramming of cellular protein production through mRNA delivered by functionalized lipid nanoparticles

2018 ◽  
Vol 115 (15) ◽  
pp. E3351-E3360 ◽  
Author(s):  
Marianna Yanez Arteta ◽  
Tomas Kjellman ◽  
Stefano Bartesaghi ◽  
Simonetta Wallin ◽  
Xiaoqiu Wu ◽  
...  
Keyword(s):  
Small Angle ◽  
Messenger Rna ◽  
Rational Design ◽  
Protein Production ◽  
Surface Composition ◽  
Cationic Lipid ◽  
Lipid Nanoparticles ◽  
Cellular Protein ◽  
Human Adipocytes ◽  
Characteristic Distance

The development of safe and efficacious gene vectors has limited greatly the potential for therapeutic treatments based on messenger RNA (mRNA). Lipid nanoparticles (LNPs) formed by an ionizable cationic lipid (here DLin-MC3-DMA), helper lipids (distearoylphosphatidylcholine, DSPC, and cholesterol), and a poly(ethylene glycol) (PEG) lipid have been identified as very promising delivery vectors of short interfering RNA (siRNA) in different clinical phases; however, delivery of high-molecular weight RNA has been proven much more demanding. Herein we elucidate the structure of hEPO modified mRNA-containing LNPs of different sizes and show how structural differences affect transfection of human adipocytes and hepatocytes, two clinically relevant cell types. Employing small-angle scattering, we demonstrate that LNPs have a disordered inverse hexagonal internal structure with a characteristic distance around 6 nm in presence of mRNA, whereas LNPs containing no mRNA do not display this structure. Furthermore, using contrast variation small-angle neutron scattering, we show that one of the lipid components, DSPC, is localized mainly at the surface of mRNA-containing LNPs. By varying LNP size and surface composition we demonstrate that both size and structure have significant influence on intracellular protein production. As an example, in both human adipocytes and hepatocytes, protein expression levels for 130 nm LNPs can differ as much as 50-fold depending on their surface characteristics, likely due to a difference in the ability of LNP fusion with the early endosome membrane. We consider these discoveries to be fundamental and opening up new possibilities for rational design of synthetic nanoscopic vehicles for mRNA delivery.

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