Tuning the Expression Level of Recombinant Proteins by Modulating MRNA Stability in a Cell-free Protein Synthesis System
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In this work, it was discovered that the stability of mRNA in a cell-free extract could be controlled by using engineered T7 terminator sequences. Specifically, it was found that mRNA stability gradually decreased as the length of the stem structure of the T7 terminator was reduced sequentially. As a result of the controlled abundance of mRNA species, it was possible to manipulate the relative expression level of target proteins by employing the T7 terminator of adjusted stem lengths.
Ren J, Zhang J, Mei Z, Shao J, Xu G, Li H Synth Syst Biotechnol. 2025; 10(1):326-335.
PMID: 39758116 PMC: 11696848. DOI: 10.1016/j.synbio.2024.11.006.
Cell-free gene expression: an expanded repertoire of applications.
Silverman A, Karim A, Jewett M Nat Rev Genet. 2019; 21(3):151-170.
PMID: 31780816 DOI: 10.1038/s41576-019-0186-3.
Cheong D, Park S, Lim H, Kim G Microorganisms. 2019; 7(1).
PMID: 30626052 PMC: 6352095. DOI: 10.3390/microorganisms7010009.
How to achieve high-level expression of microbial enzymes: strategies and perspectives.
Liu L, Yang H, Shin H, Chen R, Li J, Du G Bioengineered. 2013; 4(4):212-23.
PMID: 23686280 PMC: 3728192. DOI: 10.4161/bioe.24761.
In vitro evolution of enzymes.
Golynskiy M, Haugner 3rd J, Morelli A, Morrone D, Seelig B Methods Mol Biol. 2013; 978:73-92.
PMID: 23423890 PMC: 6924924. DOI: 10.1007/978-1-62703-293-3_6.