Effective Use of Linear DNA in Cell-Free Expression Systems
| Autor: | Megan A. McSweeney, Mark P. Styczynski |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
Histology Computer science Scientific discovery Biomedical Engineering Bioengineering Computational biology Review DNA Aptamers 01 natural sciences Expression templates 03 medical and health sciences Synthetic biology chemistry.chemical_compound genetic circuits nuclease inhibition Plasmid dna 010608 biotechnology linear expression template 030304 developmental biology rapid prototyping 0303 health sciences cell-free expression Bioengineering and Biotechnology DNA aptamers Expression (mathematics) chemistry Bacterial lysate DNA TP248.13-248.65 Biotechnology |
| Zdroj: | Frontiers in Bioengineering and Biotechnology Frontiers in Bioengineering and Biotechnology, Vol 9 (2021) |
| ISSN: | 2296-4185 |
| Popis: | Cell-free expression systems (CFEs) are cutting-edge research tools used in the investigation of biological phenomena and the engineering of novel biotechnologies. While CFEs have many benefits over in vivo protein synthesis, one particularly significant advantage is that CFEs allow for gene expression from both plasmid DNA and linear expression templates (LETs). This is an important and impactful advantage because functional LETs can be efficiently synthesized in vitro in a few hours without transformation and cloning, thus expediting genetic circuit prototyping and allowing expression of toxic genes that would be difficult to clone through standard approaches. However, native nucleases present in the crude bacterial lysate (the basis for the most affordable form of CFEs) quickly degrade LETs and limit expression yield. Motivated by the significant benefits of using LETs in lieu of plasmid templates, numerous methods to enhance their stability in lysate-based CFEs have been developed. This review describes approaches to LET stabilization used in CFEs, summarizes the advancements that have come from using LETs with these methods, and identifies future applications and development goals that are likely to be impactful to the field. Collectively, continued improvement of LET-based expression and other linear DNA tools in CFEs will help drive scientific discovery and enable a wide range of applications, from diagnostics to synthetic biology research tools. |
| Databáze: | OpenAIRE |
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