SELEX-discovered aptamer that inhibits cellular interleukin-17/interleukin-17 receptor interaction and antagonizes interleukin-17 signaling.

This research is based on a Systematic Evolution of Ligands by EXponential enrichment, also referred to as in vitro selection against the extracellular domain of human interleukin-17 receptor A (IL-17RA). Pull-down assay via quantitative polymerase chain reaction and chemiluminescence detection showed that the cloned RNA with the enriched sequence bound to human IL-17RA and inhibited the interaction between IL-17RA and human interleukin-17A (IL-17A). We also revealed that the newly discovered IL-17RA-binding RNA aptamer bound to cellular IL-17RA, inhibited the cellular IL-17RA/IL-17A interaction, and antagonized cellular IL-17A signaling.

Discovery of IL-5-binding unnatural cyclic peptides from multiple libraries by directed evolution.

Interleukin-5 (IL-5) is a type 2 cytokine involved in various allergic diseases, including severe eosinophilic asthma. In this study, we performed directed evolution against human IL-5 using systematic evolution of ligands by exponential enrichment (SELEX) from multiple mRNA-displayed peptide libraries. Peptide libraries were prepared with Escherichia coli-based reconstituted cell-free transcription and translation coupling system (PURE system) and spontaneously cyclized using multiple intramolecularly thiol-reactive benzoic acid-derived linkers, which were ribosomally incorporated through genetic code expansion. We successfully identified multiple novel IL-5-binding unnatural cyclic peptides with different cyclization linkers from multiple highly diverse mRNA-displayed libraries. Chemical dimerization was also performed to increase the avidity of unnatural cyclic IL-5-binding peptides. The novel IL-5-binding unnatural cyclic peptides discovered in this study could be used in various research, therapeutic, and diagnostic applications involving IL-5 signaling.

Artificial aptamer that inhibits interleukin-23/interleukin-23 receptor interaction discovered via SELEX.

Interaction between the pro-inflammatory cytokine interleukin-23 (IL-23) and IL-23 receptor (IL-23R) is related to the development of inflammatory autoimmune diseases such as psoriasis, inflammatory bowel disease, and Crohn's disease. In this study, we conducted systematic evolution of ligands by exponential enrichment (SELEX) for in vitro selection against human IL-23 and observed RNA sequence enrichment in the final SELEX round. IL-23-pull-down assay by chemiluminescence detection and fluorescence imaging demonstrated that SELEX-enriched RNA clone bound to IL-23. Quantitative polymerase chain reaction-based pull-down assay using the IL-23 alpha (IL-23A) subunit, a component of the IL-23 heterodimer, indicated that the RNA clone bound to IL-23A, which is favorable for autoimmune disease treatment. We also observed that the novel IL-23-binding RNA aptamer inhibited interaction between IL-23 and IL-23R. Thus, the novel IL-23-binding RNA aptamer can be used for IL-23 studies and has potential to be used for IL-23 diagnosis and IL-23-related inflammatory autoimmune disease treatment.

In vitro display evolution of unnatural peptides spontaneously cyclized via intramolecular nucleophilic aromatic substitutions.

We report novel, ribosomally incorporatable, and intramolecularly cysteine-reactive fluorobenzoic acid-derived linkers for SELEX of mRNA-displayed unnatural peptides, which spontaneously cyclized via intramolecular nucleophilic aromatic substitutions forming thioethers. With this strategy we identified several novel PCSK9-binding peptides.

Growth optimization of single-phase novel colloidal perovskite Cs3Bi2I9 nanocrystals and Cs3Bi2I9@SiO2 core-shell nanocomposites for bio-medical application.

Lead-free halide perovskites have gained attention in recent years as viable materials with more distinctive characteristics than conventional semiconductor materials. Lead-free Cs3Bi2I9 colloidal perovskite nanocrystal is chosen to eliminate its single-phase synthesis difficulty and implement the material in bioimaging applications. Nanostructured Cs3Bi2I9 perovskite composites were coated with a thin coating of SiO2 by an in situ tetraethyl orthosilicate/(3-aminopropyl)trimethoxysilane injection growth method to enhance their stability in aqueous medium and biocompatibility. Single-phase novel Cs3Bi2I9 colloidal perovskite nanocrystal synthesis was successfully developed and optimized by adopting different synthetic conditions with varied experimental parameters. Characterization studies, including X-ray diffractometry and transmission electron microscopy, confirm the hexagonal structure of Cs3Bi2I9 crystals and their cubic morphology. A broad emission peak in the red region was captured for pure and composite perovskite under different excitation wavelengths and was observed using a UV-visible spectrophotometer. Bioimaging of Cs3Bi2I9@SiO2 composites incorporated with L929 cells was conducted using an inverted fluorescence microscope under blue and green excitation. The results obtained from bioimaging studies indicated that the Cs3Bi2I9@SiO2 nanocomposites entered the cell field and exhibited an emission under excitation. The non-toxic behavior of the synthesized Cs3Bi2I9@SiO2 composites was demonstrated using MTT cytotoxicity assay in L929 fibroblast mouse cells, showing better cell compatibility.