We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
Targeted and high-throughput gene knockdown in diverse bacteria using synthetic sRNAs.
- Authors
Cho, Jae Sung; Yang, Dongsoo; Prabowo, Cindy Pricilia Surya; Ghiffary, Mohammad Rifqi; Han, Taehee; Choi, Kyeong Rok; Moon, Cheon Woo; Zhou, Hengrui; Ryu, Jae Yong; Kim, Hyun Uk; Lee, Sang Yup
- Abstract
Synthetic sRNAs allow knockdown of target genes at translational level, but have been restricted to a limited number of bacteria. Here, we report the development of a broad-host-range synthetic sRNA (BHR-sRNA) platform employing the RoxS scaffold and the Hfq chaperone from Bacillus subtilis. BHR-sRNA is tested in 16 bacterial species including commensal, probiotic, pathogenic, and industrial bacteria, with >50% of target gene knockdown achieved in 12 bacterial species. For medical applications, virulence factors in Staphylococcus epidermidis and Klebsiella pneumoniae are knocked down to mitigate their virulence-associated phenotypes. For metabolic engineering applications, high performance Corynebacterium glutamicum strains capable of producing valerolactam (bulk chemical) and methyl anthranilate (fine chemical) are developed by combinatorial knockdown of target genes. A genome-scale sRNA library covering 2959 C. glutamicum genes is constructed for high-throughput colorimetric screening of indigoidine (natural colorant) overproducers. The BHR-sRNA platform will expedite engineering of diverse bacteria of both industrial and medical interest. Using synthetic sRNAs to knockdown target genes has been restricted to a limited number of bacteria. Here, the authors develop a broad-host-range synthetic sRNA platform and show its application in 16 bacterial species, including mitigating virulence-associated phenotypes in pathogens and production of chemicals via metabolic engineering.
- Subjects
GENE targeting; CORYNEBACTERIUM glutamicum; BACTERIA; STAPHYLOCOCCUS epidermidis; KLEBSIELLA pneumoniae; HIGH throughput screening (Drug development)
- Publication
Nature Communications, 2023, Vol 14, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-023-38119-y