Establishment of a Secretory Protein-Inducible CRISPR/Cas9 System for Nosema bombycis in Insect Cells.

Gene editing techniques are widely and effectively used for the control of pathogens, but it is difficult to directly edit the genes of Microsporidia due to its unique spore wall structure. Innovative technologies and methods are urgently needed to break through this limitation of microsporidia therapies. Here, we establish a microsporidia-inducible gene editing system through core components of microsporidia secreted proteins, which could edit target genes after infection with microsporidia. We identified that Nosema bombycis NB29 is a secretory protein and found to interact with itself. The NB29-N3, which lacked the nuclear localization signal, was localized in the cytoplasm, and could be tracked into the nucleus after interacting with NB29-B. Furthermore, the gene editing system was constructed with the Cas9 protein expressed in fusion with the NB29-N3. The system could edit the exogenous gene EGFP and the endogenous gene BmRpn3 after overexpression of NB29 or infection with N. bombycis.

Enhancement of peroxymonosulfate activation for 2,4-dichlorophenoxyacetic acid removal by MoSe2 induced Fe redox cycles.

The limited regeneration of Fe2+ in the Fe-catalyzed advanced oxidation processes (AOPs) constrained its application for the removal of organic pollutants. Herein, MoSe2 was introduced to promote the degradation of 2,4-dichlorophenoxyacetic acid (2,4-D) in the Fe2+/PMS system. Compared with Fe2+/PMS processes, the 2,4-D degradation efficiency and PMS decomposition rate respectively increased by 73.8% and 84.2% in the MoSe2/Fe2+/PMS system. DFT simulation results suggested that Se atoms acted smoothly as the bridge supporting the charge transfer from Mo to adjacent Fe atoms, which led to the reduction of Fe3+. The rapid regeneration of Fe2+ boosted the activation of PMS and the degradation of pollutants. Additionally, the electron paramagnetic resonance (EPR) and quenching experiments results indicated that SO4∙-, ∙OH, and 1O2 accounted for 2,4-D degradation, and SO4∙- and 1O2 predominated the reaction. The Mo based co-catalysts showed better co-catalytic effect than the W counterparts, and the moderate adsorption for PMS and lower electron transfer electron transfer resistance accounted for the more excellent co-catalytic performance of MoSe2 than that of WSe2. In addition, the degradation efficiency of 2,4-D was up to 95.5% after five cycles of MoSe2 in the co-catalytic system. The coexistent humic acid (HA) and Cl- showed ignorant negative effect on the degradation, while HCO3- would depress the oxidation reaction. The acidic etching wastewater can be applied as the Fe ions source in this co-catalytic process to remove 2,4-D effectively.