• MeSH
• Oligonucleotides, Antisense physiology   MeSH
• Gene Expression MeSH
• Lipids MeSH
• Genes, Viral MeSH
• Hepatitis B virus genetics   growth & development   MeSH
• Publication type
• Comparative Study MeSH

• MeSH
• Antisense Elements (Genetics) MeSH
• Oligonucleotides, Antisense MeSH
• Gene Expression drug effects   MeSH
• Nucleic Acids, Nucleotides, and Nucleosides MeSH

• Conspectus
• Biochemie. Molekulární biologie. Biofyzika
• NML Fields
• genetika, lékařská genetika
• biochemie
• biologie
• NML Publication type
• elektronické časopisy

Effective translation of rare disease diagnosis knowledge into therapeutic applications is achievable within a reasonable timeframe; where mutations are amenable to current antisense oligonucleotide technology. In our study, we identified five distinct types of abnormal splice-causing mutations in patients with rare genetic disorders and developed a tailored antisense oligonucleotide for each mutation type using phosphorodiamidate morpholino oligomers with or without octa-guanidine dendrimers and 2'-O-methoxyethyl phosphorothioate. We observed variations in treatment effects and efficiencies, influenced by both the chosen chemistry and the specific nature of the aberrant splicing patterns targeted for correction. Our study demonstrated the successful correction of all five different types of aberrant splicing. Our findings reveal that effective correction of aberrant splicing can depend on altering the chemical composition of oligonucleotides and suggest a fast, efficient, and feasible approach for developing personalized therapeutic interventions for genetic disorders within short time frames.

• MeSH
• Oligonucleotides, Antisense * therapeutic use   genetics   MeSH
• Genetic Diseases, Inborn genetics   therapy   MeSH
• Humans MeSH
• Morpholinos therapeutic use   genetics   MeSH
• Mutation * MeSH
• RNA Splicing * MeSH
• Rare Diseases * genetics   drug therapy   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

• MeSH
• Adenosine Monophosphate chemistry   MeSH
• Oligonucleotides, Antisense chemistry   metabolism   MeSH
• Nucleic Acid Hybridization MeSH
• Organophosphonates chemistry   MeSH

• MeSH
• Antisense Elements (Genetics) MeSH
• Gene Expression drug effects   MeSH
• Pharmacology MeSH

• Conspectus
• Biochemie. Molekulární biologie. Biofyzika
• NML Fields
• genetika, lékařská genetika
• biologie
• NML Publication type
• elektronické časopisy

Previous reports from this laboratory have shown marked cytocidal effects of the ISIS-3466 antisense phosphorothioate oligodeoxynucleotide to the human nucleolar protein p120 on human cancer cell lines in vitro and inhibition of tumor growth in vivo in an i.p/i.p. LOX cell model (L. Perlaky et al. Anti-Cancer Drug Design 8:3-14, 1993). In this study, light and fluorescence microscopy showed that the number of LOX cells in mitosis decreased by 50% after incubation for 4 h in 0.2-0.4 microM antisense oligonucleotide; a 70% reduction in cell number was found from 8-72 h post-treatment. In addition, marked unravelling of nucleolar structures and chromatin fragmentation was found after a 4-h incubation. The nucleolar unravelling occurred in varying degrees ranging from partial unfolding to almost complete separation of the strands of nucleolar residues. Twenty four hours post-treatment, immunofluorescence staining with the anti-p120 monoclonal antibody showed reduced nucleolar protein p120 and translocation of the p120 protein from the nucleoli to the nucleoplasm. Analysis of the mechanisms of the nucleolar unravelling and inhibition of mitosis will provide further understanding of the cytocidal effects of the ISIS-3466 antisense oligonucleotide.

• MeSH
• Melanoma, Amelanotic * genetics   chemistry   pathology   MeSH
• Antigens, Neoplasm * drug effects   MeSH
• Oligodeoxyribonucleotides, Antisense * MeSH
• Oligonucleotides, Antisense pharmacology   genetics   MeSH
• Cell Nucleolus * drug effects   MeSH
• Cell Nucleus drug effects   MeSH
• Chromatin drug effects   MeSH
• Microscopy, Fluorescence MeSH
• Nuclear Proteins * drug effects   MeSH
• Humans MeSH
• Mitosis drug effects   MeSH
• Molecular Sequence Data MeSH
• Tumor Cells, Cultured drug effects   MeSH
• Base Sequence MeSH
• Thionucleotides pharmacology   genetics   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH

BACKGROUND: The genetic and epigenetic alterations observed in acute myeloid leukemia (AML) contribute to its heterogeneity, influencing disease progression response to therapy, and patient outcomes. The use of antisense oligonucleotides (ASOs) technology allows for the design of oligonucleotide inhibitors based on gene sequence information alone, enabling precise targeting of key molecular pathways or specific genes implicated in AML. METHODS AND RESULTS: Midostaurin, a FLT3 specific inhibitor and ASOs targeting particular genes, exons, or mutations was conducted using AML models. This ASOs treatment was designed to bind to exon 7 of the MBNL1 (muscleblind-like) gene. Another target was the FLT3 gene, focusing on two aspects: (a) FLT3-ITD (internal tandem duplication), to inhibit the expression of this aberrant gene form, and (b) the FLT3 in general. Treated and untreated cells were analyzed using quantitative PCR (qPCR), dot blot, and Raman spectroscopy. This study contrasts midostaurin with ASOs that inhibit FLT3 protein production or its isoforms via mRNA degradation. A trend of increased FLT3 expression was observed in midostaurin-treated cells, while ASO-treated cells showed decreased expression, though these changes were not statistically significant. CONCLUSIONS: In AML, exon 7 of MBNL1 is involved in several cellular processes and in this study, exon 7 of MBNL1 was targeted for method optimization, with the highest block of the exon 7 gene variant observed 48 h post-transfection. Midostaurin, a multitargeted kinase inhibitor, acts against the receptor tyrosine kinase FLT3, a critical molecule in AML pathogenesis. While midostaurin blocks FLT3 signaling pathways, it paradoxically increases FLT3 expression.

• MeSH
• Leukemia, Myeloid, Acute * genetics   drug therapy   MeSH
• Oligonucleotides, Antisense * pharmacology   genetics   MeSH
• Exons genetics   MeSH
• Humans MeSH
• Cell Line, Tumor MeSH
• RNA-Binding Proteins genetics   metabolism   MeSH
• Gene Expression Regulation, Leukemic drug effects   MeSH
• Staurosporine * analogs & derivatives   pharmacology   MeSH
• fms-Like Tyrosine Kinase 3 * genetics   antagonists & inhibitors   metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

• MeSH
• Oligonucleotides, Antisense chemistry   therapeutic use   MeSH
• Cytidine chemistry   MeSH
• Phosphates chemistry   MeSH
• Guanosine chemistry   MeSH
• Models, Molecular MeSH
• Binding Sites analysis   chemistry   MeSH

• MeSH
• Acquired Immunodeficiency Syndrome drug therapy   MeSH
• Oligonucleotides, Antisense MeSH
• HIV drug effects   MeSH
• RNA, Catalytic pharmacology   MeSH
• RNA, Viral drug effects   MeSH
• Zidovudine pharmacology   MeSH
• Zinc Fingers antagonists & inhibitors   MeSH

Specific gene knockdown mediated by the antisense oligodeoxynucleotides (AODNs) strategy recently emerged as a rapid and effective tool for probing gene role in plant cells, particularly tip-growing pollen tubes. Here, we describe the protocol for the successful employment of AODN technique in growing tobacco pollen tubes, covering AODN design, application, and analysis of the results. We also discuss the advantages and drawbacks of this method.

• MeSH
• Oligonucleotides, Antisense genetics   MeSH
• Phenotype MeSH
• Gene Knockdown Techniques * MeSH
• Pollen Tube genetics   growth & development   MeSH
• Gene Expression Regulation, Plant MeSH
• Nicotiana genetics   growth & development   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH