Elucidation of gene function using C-5 propyne antisense oligonucleotides
| Autor: | Flanagan Wm, Leon Su, Wagner Rw |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Biomedical Engineering
Gene Expression Breast Neoplasms Bioengineering Cyclin B Biology Applied Microbiology and Biotechnology CDC2 Protein Kinase Tumor Cells Cultured Humans RNA Messenger RNA Neoplasm Cyclin B1 Cell Cycle Protein Mitosis Cyclin Base Sequence Cell Cycle Oligonucleotides Antisense Thionucleotides Cell cycle Molecular biology Cell biology Antisense RNA Cell culture Gene Targeting Cancer cell Molecular Medicine Female Cell Division Biotechnology |
| Zdroj: | Nature Biotechnology. 14:1139-1145 |
| ISSN: | 1546-1696 1087-0156 |
| DOI: | 10.1038/nbt0996-1139 |
| Popis: | Identification of human disease-causing genes continues to be an intense area of research. While cloning of genes may lead to diagnostic tests, development of a cure requires an understanding of the gene's function in both normal and diseased cells. Thus, there exists a need for a reproducible and simple method to elucidate gene function. We evaluate C-5 propyne pyrimidine modified phosphorothioate antisense oligonucleotides (ONs) targeted against two human cell cycle proteins that are aberrantly expressed in breast cancer: p34cdc2 kinase and cyclin B1. Dose-dependent, sequence-specific, and gene-specific inhibition of both proteins was achieved at nanomolar concentrations of ONs in normal and breast cancer cells. Precise binding of the antisense ONs to their target RNA was absolutely required for antisense activity. Four or six base-mismatched ONs eliminated antisense activity confirming the sequence specificity of the antisense ONs. Antisense inhibition of p34cdc2 kinase resulted in a significant accumulation of cells in the Gap2/mitosis phase of the cell cycle in normal cells, but caused little effect on cell cycle progression in breast cancer cells. These data demonstrate the potency, specificity, and utility of C-5 propyne modified antisense ONs as biological tools and illustrate the redundancy of cell cycle protein function that can occur in cancer cells. |
| Databáze: | OpenAIRE |
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