Broad-Spectrum Gene Repression Using Scaffold Engineering of Synthetic sRNAs
- Authors
- Noh, Minho; Yoo, Seung Min; Yang, Dongsoo; Lee, Sang Yup
- Issue Date
- Jun-2019
- Publisher
- AMER CHEMICAL SOC
- Keywords
- gene repression; Hfq-binding region; knockdown; synthetic sRNA
- Citation
- ACS SYNTHETIC BIOLOGY, v.8, no.6, pp 1452 - 1461
- Pages
- 10
- Journal Title
- ACS SYNTHETIC BIOLOGY
- Volume
- 8
- Number
- 6
- Start Page
- 1452
- End Page
- 1461
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/32748
- DOI
- 10.1021/acssynbio.9b00165
- ISSN
- 2161-5063
- Abstract
- Gene expression regulation in broad-spectrum range is critical for constructing cell factories and genetic circuits to balance and control system-wide fluxes. Synthetic small regulatory RNAs (sRNAs) effectively regulate gene expression at the translational level by modulating an mRNA-binding chance and sRNA abundance; however, it can control target gene expression only within the limit of the intrinsic repression ability of sRNAs. Here, we systematically mutated a SgrS scaffold as a model sRNA by dividing the Hfq-binding module of the sRNA into the three regions: the A/U-rich sequence, the stem, and the hairpin loop, and examined how efficiently the mutants suppressed DsRed2 expression. By doing this, we found that a scaffold with an altered A/U-rich sequence (CUUU) and stem length and that with altered A/U-rich sequence (GCAC) showed a 3-fold stronger and a 3-fold weaker repression than the original scaffold, respectively. For practical application of altered scaffolds, proof-of-concept experiments were performed by constructing a library of 67 synthetic sRNAs with the strongest scaffold, each one targeting a different rationally selected gene, and using this library to enhance cadaverine production in Escherichia coli, yielding in 27% increase (1.67 g/L in flask cultivation, 13.7 g/L in fed-batch cultivation). Synthetic sRNAs with engineered sRNA scaffolds could be useful in modulating gene expression for strain improvement.
- Files in This Item
- There are no files associated with this item.
- Appears in
Collections - College of ICT Engineering > School of Integrative Engineering > 1. Journal Articles
![qrcode](https://api.qrserver.com/v1/create-qr-code/?size=55x55&data=https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/32748)
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.