Spinal Muscular Atrophy (SMA) is a fatal neuromuscular disease characterized by motor neuron loss and advanced muscle weakness, which occurs in functional SMN (Survival Motor Neuron) protein deficiency with SMN1 gene-induced deletions and mutations. The incidence of SMA, which is an autosomal recessive disease, is 1/10,000 in the world. The SMN protein acts as a molecular chaperone in the formation of the spliceosome complex, which catalyzes the splicing of pre-mRNA, enabling mRNAs and non-coding RNAs to mature. Since the current SMN1 -encoding Adeno-associated virus (AAV) or SMN2 gene targeting antisense oligonucleotide-based strategies cannot provide long-term stable SMN expression in neuron cells, more effective methods need to be developed. CRISPR technology, which adds a new dimension to genetic engineering and gene therapies, makes it possible to treat many genetic diseases. In terms of SMA, some previous studies in the literature prove that it is possible to treat SMA with the CRISPR strategy. Homology Directed Repair (HDR)-based CRISPR technology, which results in a high rate of in-del (insertion-deletion) mutations rather than editing, was shown unsuitable for therapeutic applications. CRISPR-Prime editing (PE) technology is a new generation of gene editing approach that precisely provides various genomic modifications without the need for double-strand breakage or donor DNA sequences. CRISPR-Prime Editing method has also been used in rare diseases such as sickle cell anemia and Tay-Sachs, and their efficiency in editing various pathogenic mutations has been demonstrated. However, CRISPR Prime Editing-mediated gene editing for Spinal Muscular Atrophy (SMA) have not yet been investigated. The c.840 T-C transition and c.859 G-C transformations in the SMN2 gene and the correction of these point mutations with a single pegRNA at the same time were investigated for the first time in this study. Here, we showed that CRISPR-PE systems could increase SMN2 gene activity and SMN protein expression by ensuring exon 7 participation by editing c.840 T-C transition and c.859 G-C transformations. The fact that Prime Editing method showed the efficacy and stability of modifications in SMN2 genes that were investigated in SMN-low Jurkat cells as a proof-of-concept. This study enabled the next step with the CRISPR-Prime Editing approach to be tested ex vivo in primary cell lines from SMA patients and SMN-low neuronal cells.

Carvacrol, a phenolic monoterpene predominantly found in Origanum species, has been reported to exhibit antimicrobial, anti-inflammatory, antioxidant, and cytotoxic effects. Formulations such as Vacrol and S-Mix, enriched with carvacrol and complementary essential oil compounds, may enhance therapeutic efficacy while reducing toxicity. Essential oil components were analyzed via GC-MS. Cell viability was assessed using the sulforhodamine B (SRB) assay at different concentrations and incubation periods. An in ovo chorioallantoic membrane (CAM) assay was performed to investigate tumor volume changes and histopathological alterations. Vacrol and S-Mix demonstrated concentration- and time-dependent cell viability-attenuating effects in MDA-MB-231 cells, with significant reductions in viability at higher concentrations (1–10 mM). In ovo, S-Mix induced ~40% reduction in tumor volume and promoted apoptotic morphology compared to controls. Combined effects of carvacrol with α-pinene, eugenol, and β-terpineol likely contributed to enhanced bioactivity. These findings support further preclinical and mechanistic investigations to validate their therapeutic potential.

Lung cancer remains a leading cause of cancer-related mortality and is frequently complicated by respiratory infections, supporting interest in agents with both antitumoral and antimicrobial potential. This study evaluated two standardized thyme-derived, carvacrol-based formulations, Vacrol and S-Mix, in lung cancer-associated experimental models. A549 lung adenocarcinoma and BEAS-2B bronchial epithelial cells were treated with the formulations, and cell viability, clonogenic capacity, SIRT1–SIRT7 protein expression, in ovo tumor growth, histopathological changes, and antimicrobial activity against pneumonia-associated reference strains were assessed. S-Mix showed stronger short-term cytotoxicity in A549 cells, reaching an IC₅₀ of 1 mM after 72 h, whereas Vacrol produced more pronounced modulation of selected sirtuin proteins, particularly SIRT1, SIRT4, and SIRT5. Both formulations suppressed colony formation under prolonged exposure. In the CAM model, Vacrol was associated with greater macroscopic suppression of tumor growth and vascularization, while S-Mix produced more prominent histopathological cellular injury. Vacrol also showed antimicrobial activity against tested respiratory pathogens, with MIC values ranging from 0.5 to 4 mg/mL, MBC values ranging from 1 to 4 mg/mL and volatile-phase activity against Streptococcus pneumoniae as well as Klebsiella pneumoniae. These findings suggest that carvacrol-based formulations exert distinct cytotoxic, sirtuin-modulatory, antitumoral, and antimicrobial effects, warranting further mechanistic and translational validation.

Carvacrol, a phenolic monoterpene predominantly found in Origanum species, has been reported to exhibit antimicrobial, anti-inflammatory, antioxidant, and anticancer effects. Synergistic formulations such as Vacrol and S-Mix, enriched with carvacrol and complementary essential oil compounds, may enhance therapeutic efficacy while reducing toxicity. Essential oil components were analyzed via GC-MS. Cell viability was assessed using the sulforhodamine B (SRB) assay at different concentrations and incubation periods. An in ovo chorioallantoic membrane (CAM) assay was performed to investigate tumor volume changes and histopathological alterations. Vacrol and S-Mix demonstrated concentration- and time-dependent cytotoxic effects in MDA-MB-231 cells, with significant reductions in viability at higher concentrations (100 µM–1 mM). In ovo, S-Mix induced ~40% reduction in tumor volume and promoted apoptotic morphology compared to controls. Synergistic effects of carvacrol with α-pinene, eugenol, and β-terpineol likely contributed to enhanced bioactivity. Vacrol and S-Mix exhibit promising antiproliferative and pro-apoptotic activity against triple-negative breast cancer models. These findings support further preclinical and mechanistic investigations to validate their therapeutic potential.