Increased gene translation stringency in mammalian cells by nonsense suppression at multiple permissive sites with a single noncanonical amino acid
| Autor: | Iva Hafner Bratkovic, Roman Jerala, Maja Svetličič, Lucija Kadunc, Vida Forstnerič |
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| Rok vydání: | 2020 |
| Předmět: |
endocrine system diseases
Stringent response media_common.quotation_subject Phenylalanine Nonsense Biophysics Biochemistry Amino Acyl-tRNA Synthetases 03 medical and health sciences chemistry.chemical_compound Benzophenones RNA Transfer Structural Biology Gene expression Genetics Humans Molecular Biology Gene 030304 developmental biology media_common chemistry.chemical_classification 0303 health sciences Chemistry Aminoacyl tRNA synthetase 030302 biochemistry & molecular biology Translation (biology) Cell Biology Cell biology Amino acid HEK293 Cells Codon Nonsense Protein Biosynthesis Transfer RNA |
| Zdroj: | FEBS lettersReferences. 594(15) |
| ISSN: | 1873-3468 |
| Popis: | The considerable potential of engineered cells compels the development of strategies for the stringent control of gene expression. A promising approach is the introduction of a premature stop codon (PTC) into a selected gene that is expressed only in the presence of noncanonical amino acids through nonsense suppression. Here, different strategies of amber PTC readthrough in mammalian cells were tested. The use of a tRNA synthetase together with a TAG codon-specific tRNA achieved PTC readthrough depending on the addition of a noncanonical amino acid (4-benzoyl-L-phenylalanine; Bpa). While single TAG codon incorporation exhibited detectable expression of the reporter protein even in the absence of Bpa, the use of a double PTC enabled virtually leakage-free functional gene translation. The introduction of an additional 5'-PTC, therefore, represents a generally applicable strategy to increase stringency in gene translation. |
| Databáze: | OpenAIRE |
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