Development of multiplexing gene silencing system using conditionally induced polycistronic synthetic antisense RNAs in Escherichia coli

Fujita, Shouta; Tsumori, Yutaka; Makino, Yuko; Saito, Mineki; Kawano, Mitsuoki

doi:10.1016/j.bbrc.2021.03.152

Cited by 1 publication

(1 citation statement)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We first examined whether the binding of dCas13 indeed mediated the knockdown of targeted mRNA since only the formation of RNA duplex could knock down the mRNA as in the case of antisense RNA [20][21][22] . We tested various concentrations of aTc, thereby differing the expression level of dCas13, to evaluate the correlation between the intracellular amount of dCas13 and the target gene expression.…”

Section: Design and Characterization Of The Dcas13-based Tl-crispri S...mentioning

confidence: 99%

Tunable translation-level CRISPR interference by dCas13 and engineered gRNA in bacteria

Kim,

Kim

et al. 2024

Nat Commun

View full text Add to dashboard Cite

Although CRISPR-dCas13, the RNA-guided RNA-binding protein, was recently exploited as a translation-level gene expression modulator, it has still been difficult to precisely control the level due to the lack of detailed characterization. Here, we develop a synthetic tunable translation-level CRISPR interference (Tl-CRISPRi) system based on the engineered guide RNAs that enable precise and predictable down-regulation of mRNA translation. First, we optimize the Tl-CRISPRi system for specific and multiplexed repression of genes at the translation level. We also show that the Tl-CRISPRi system is more suitable for independently regulating each gene in a polycistronic operon than the transcription-level CRISPRi (Tx-CRISPRi) system. We further engineer the handle structure of guide RNA for tunable and predictable repression of various genes in Escherichia coli and Vibrio natriegens. This tunable Tl-CRISPRi system is applied to increase the production of 3-hydroxypropionic acid (3-HP) by 14.2-fold via redirecting the metabolic flux, indicating the usefulness of this system for the flux optimization in the microbial cell factories based on the RNA-targeting machinery.

show abstract

Section: Design and Characterization Of The Dcas13-based Tl-crispri S...mentioning

confidence: 99%

Tunable translation-level CRISPR interference by dCas13 and engineered gRNA in bacteria

Kim,

Kim

et al. 2024

Nat Commun

View full text Add to dashboard Cite

show abstract

Development of multiplexing gene silencing system using conditionally induced polycistronic synthetic antisense RNAs in Escherichia coli

Cited by 1 publication

References 21 publications

Tunable translation-level CRISPR interference by dCas13 and engineered gRNA in bacteria

Tunable translation-level CRISPR interference by dCas13 and engineered gRNA in bacteria

Contact Info

Product

Resources

About