No abstract

Usher syndrome (USH) is the most common form of deaf-blindness in humans. Molecular characterization revealed that the USH gene products form a macromolecular protein network in hair cells of the inner ear and in photoreceptor cells of the retina via binding to PDZ domains in the scaffold protein harmonin encoded by the Ush1c gene in mice and humans. Although several mouse mutants for the Ush1c gene have been described, we generated a targeted null mutation Ush1c mouse model in which the first four exons of the Ush1c gene were replaced with a reporter gene. Here, we assessed the expression pattern of the reporter gene under control of Ush1c regulatory elements and characterized the phenotype of mice defective for Ush1c. These Ush1 knockout mice are deaf but do not recapitulate vision defects before 10 months of age. Our data show LacZ expression in multiple layers of the retina but in neither outer nor inner segments of the photoreceptor layers in mice bearing the knockout construct at 1-5 months of age. The fact that Ush1c expression is much higher in the ear than in the eye suggests a different role for Ush1c in ear function than in the eye and may explain why Ush1c mutant mice do not recapitulate vision defects. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. KeywordsAuthors' contributions: CT performed the immunoassays. XZL contributed to initial research design. FC and CT performed the molecular studies including real-time PCR and sequence analyses. HY performed ABR tests, the immunoassays, contributed to generation of backcross and Ush1c −/− mice. CT and CLG contributed the lacZ reporter gene expression and hair bundle assay, CLG performed the SEM assay. BY and CL performed genotyping, DY performed literature research. QYZ conceived and designed the study, supervised all the experimental data analyses and the manuscript preparation. All authors read and approved the final manuscript.NIH Public Access

Recently, lipid nanoparticles (LNPs) have attracted attention due to their emergent use for COVID‐19 mRNA vaccines. The success of LNPs can be attributed to ionizable lipids, which enable functional intracellular delivery. Previously, the authors established an automated high‐throughput platform to screen ionizable lipids and identified that the LNPs generated using this automated technique show comparable or increased mRNA functional delivery in vitro as compared to LNPs prepared using traditional microfluidics techniques. In this study, the authors choose one benchmark lipid, DLin‐MC3‐DMA (MC3), and investigate whether the automated formulation technique can enhance mRNA functional delivery in vivo. Interestingly, a 4.5‐fold improvement in mRNA functional delivery in vivo by automated LNPs as compared to LNPs formulated by conventional microfluidics techniques, is observed. Mechanistic studies reveal that particles with large size accommodate more mRNA per LNP, possess more hydrophobic surface, are more hemolytic, bind a larger protein corona, and tend to accumulate more in macropinocytosomes, which may quantitatively benefit mRNA cytosolic delivery. These data suggest that mRNA loading per particle is a critical factor that accounts for the enhanced mRNA functional delivery of automated LNPs. These mechanistic findings provide valuable insight underlying the enhanced mRNA functional delivery to accelerate future mRNA LNP product development.

Herein, MoS2‐embedded human serum albumin hollow nanocapsules (denoted MoS2/HSA) are successfully prepared via a hard‐core‐assisted layer‐by‐layer coating approach. The hollow MoS2/HSA nanocapsules possess an uniform size (280 nm), a larger hollow cavity, a low Young's modulus (222 ± 20 MPa), excellent photothermal conversion ability, and a high drug loading capacity for doxorubicin (27 wt%). The flexible hollow MoS2/HSA nanocapsules exhibit significantly enhanced cellular uptake efficiency against breast cancer cells duo to their flexible property and targeting ability of HSA. In vivo experiments show that the flexible MoS2/HSA could reduce phagocytosis and effectively improve blood persistence and tumor accumulation. In vitro and in vivo experiments have demonstrated that the flexible MoS2/HSA based nanoplatforms effectively ablate breast cancer cells via synergistic photothermal and chemical therapy.