Cell-Free Transcription-Coupled CRISPR/Cas12a Assay for Prototyping Cyanobacterial Promoters
SCIE
SCOPUS
- Title
- Cell-Free Transcription-Coupled CRISPR/Cas12a Assay for Prototyping Cyanobacterial Promoters
- Authors
- Choi, Yun-Nam; Shin, Ye Rim; Park, Jong Moon; Lee, Jeong Wook
- Date Issued
- 2021-06
- Publisher
- AMER CHEMICAL SOC
- Abstract
- Cyanobacteria are promising microbial hosts for the production of diverse biofuels and biochemicals. However, compared to other model microbial hosts such as Escherichia coli and yeast, it takes a long time to genetically modify cyanobacteria. One way to efficiently engineer cyanobacteria while minimizing genetic engineering would be to develop a fast, high-throughput prototyping tool for cyanobacteria. In this study, we developed a CRISPR/Cas12a-based assay coupled with cyanobacteria cell-free systems to rapidly prototype promoter characteristics. Using this newly developed assay, we demonstrated cyanobacteria cell-free transcription for the first time and confirmed a positive correlation between the in vitro and in vivo transcription performance. Furthermore, we generated a synthetic promoter library and evaluated the characteristics of promoter subregions by using the assay. Varied promoter strength derived from random mutations were rapidly and effectively measured in a high-throughput way. We believe that this study offers an easily applicable and rapid prototyping platform to characterize promoters for cyanobacterial engineering.
- Keywords
- article; cell free system; CRISPR Cas system; cyanobacterium; in vitro study; in vivo study; nonhuman; promoter region; rapid prototyping
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/106820
- DOI
- 10.1021/acssynbio.1c00148
- ISSN
- 2161-5063
- Article Type
- Article
- Citation
- ACS SYNTHETIC BIOLOGY, vol. 10, no. 6, page. 1300 - 1307, 2021-06
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- There are no files associated with this item.
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