Small interfering RNA (siRNA) and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated Protein 9 (Cas9) complexes are effective approaches to temporarily downregulate protein expression via post-transcription RNA interference or permanently altering protein expression via editing genomic DNA, respectively. However, the efficient delivery of these mediators to targeted cells has been challenging, largely due to their anionic and hydrophilic nature, which hinders their interaction with the cell membrane and cellular internalization. Cell-penetrating peptides (CPPs) exhibit dual characteristics as a carrier for nucleic acid delivery, where positively charged components bind to the negatively charged nucleic acid, and the hydrophobic components enhance interactions with the cell membranes. Arginine/tryptophan-containing peptides provide both characteristics and have been shown to be efficient in delivering nucleic acids.
In this project, we evaluated a library of novel linear and hybrid (cyclic/linear) peptides with increasing molecular weight for their physical characteristics, interaction with nucleic acids, cytotoxicity, internalization into different breast cancer cell lines, and their ability to promote and enhance silencing efficiency.
The majority of the peptides included in this study demonstrated strong binding affinity to siRNA; however, despite effective cellular internalization, they did not show expected silencing/transfection efficiency, which we hypothesized to be due to the strong bonding between the carrier and cargo and, therefore, poor intracellular release of delivered nucleic acids.
The addition of a negatively charged component to the peptide/siRNA and peptide/ribonucleoprotein complexes enhanced the efficiency of the delivered nucleic acid. These data emphasize the importance of the balance between stability of the carrier/cargo complex and timely intracellular release in the efficiency of this approach.
Abdulelah Alhazza, Sorour Khayyatnejad Shoushtari, Hasan Uludağ et al.· Frontiers in Nanotechnology· 0 citations
Rheumatoid arthritis (RA) is a chronic immune-mediated disease in which persistent inflammation and oxidative stress contribute to progressive joint damage. This study evaluated the prophylactic effects of sabinene in complete Freund's adjuvant (CFA)-induced arthritis and explored potential molecular associations using integrated in vivo and computational approaches. Rats were allocated to vehicle control, arthritic control, piroxicam, or sabinene groups (15, 30, or 60 mg/kg; n = 6/group); sabinene was administered orally beginning 30 minutes before CFA injection and continued for 28 days. Paw swelling, hematological and biochemical parameters, inflammatory and oxidative-stress markers, and gene expression were assessed, followed by network pharmacology, enrichment analysis, and molecular docking. Sabinene dose-dependently attenuated CFA-induced paw swelling and body-weight loss and improved hematological, hepatic, renal, and inflammatory parameters. Treatment also enhanced antioxidant defenses and reduced lipid peroxidation, prostaglandin E₂, 5-lipoxygenase, and anti-cyclic citrullinated peptide antibody levels. Sabinene reduced messenger RNA expression of nuclear factor kappa B (NF-κB), toll-like receptor 4 (TLR4), NOD-like receptor family pyrin domain-containing 3 (NLRP3), caspase-1, gasdermin D, and other pro-inflammatory genes while increasing interleukin-4 and interleukin-10 expression. Network pharmacology identified 133 overlapping sabinene-RA targets, and molecular docking predicted interactions with tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), signal transducer and activator of transcription 3 (STAT3), and interferon gamma (IFN-γ). These findings support a prophylactic protective effect of sabinene against CFA-induced arthritis associated with reduced inflammation, improved antioxidant status, and modulation of inflammasome-related gene expression. Further protein-level, functional, and post-induction studies are required to confirm the proposed mechanisms and therapeutic relevance.
Kanwal Asif, Ambreen Malik Uttra, Arham Shabbir et al.· Biomolecules & biomedicine· 0 citations
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