Synthetic fused sRNA for the simultaneous repression of multiple genes.

Efficient control over multiple gene expression still presents a major challenge. Synthetic sRNA enables targeted gene expression control in trans without directly modifying the chromosome, but its use to simultaneously target multiple genes can often cause cell growth defects because of the need for additional energy for transcription and lowering of their repression efficiency by limiting the amount of Hfq protein. To address these limitations, we present fusion sRNA (fsRNA) that simultaneously regulates the translation of multiple genes efficiently. It is constructed by linking the mRNA-binding modules for multiple targeted genes in one sRNA scaffold via one-pot generation using overlap extension PCR. The repression capacity of fsRNA was demonstrated by the construction of sRNAs to target four endogenous genes: caiF, hybG, ytfR and minD in Escherichia coli. Their cross-reactivity and the effect on cell growth were also investigated. As practical applications, we applied fsRNA to violacein- and protocatechuic acid-producing strains, resulting in increases of 13% violacein and 81% protocatechuic acid, respectively. The developed fsRNA-mediated multiple gene expression regulation system thus enables rapid and efficient development of optimised cell factories for valuable chemicals without cell growth defects and limiting cellular resources.Key points• Synthetic fusion sRNA (fsRNA)-based system was constructed for the repression of multiple target genes.• fsRNA repressed multiple genes by only expressing a single sRNA while minimising the cellular burden.• The application of fsRNA showed the increased production titers of violacein (13%) and protocatechuic acid (81%).

Electrochemiluminescence Systems for the Detection of Biomarkers: Strategical and Technological Advances.

Electrochemiluminescence (ECL)-based sensing systems rely on light emissions from luminophores, which are generated by high-energy electron transfer reactions between electrogenerated species on an electrode. ECL systems have been widely used in the detection and monitoring of diverse, disease-related biomarkers due to their high selectivity and fast response times, as well as their spatial and temporal control of luminance, high controllability, and a wide detection range. This review focuses on the recent strategic and technological advances in ECL-based biomarker detection systems. We introduce several sensing systems for medical applications that are classified according to the reactions that drive ECL signal emissions. We also provide recent examples of sensing strategies and technologies based on factors that enhance sensitivity and multiplexing abilities as well as simplify sensing procedures. This review also discusses the potential strategies and technologies for the development of ECL systems with an enhanced detection ability.