Polyamide fluorescent probes for visualization of repeated DNA sequences in living cells
| Autor: | Karine Nozeret, Alexandre S. Boutorine, Christophe Escudé, François Loll |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
DNA nanoball sequencing
Circular dichroism Satellite DNA Biology Biochemistry Genome Fluorescence spectroscopy DNA sequencing chemistry.chemical_compound Mice Animals Molecular Biology Fluorescent Dyes Repetitive Sequences Nucleic Acid Base Sequence Organic Chemistry Optical Imaging DNA Fluorescence Nylons chemistry Microscopy Fluorescence NIH 3T3 Cells Molecular Medicine |
| Zdroj: | Chembiochem : a European journal of chemical biology. 16(4) |
| ISSN: | 1439-7633 |
| Popis: | DNA imaging in living cells usually requires transgenic approaches that modify the genome. Synthetic pyrrole-imidazole polyamides that bind specifically to the minor groove of double-stranded DNA (dsDNA) represent an attractive approach for in-cell imaging that does not necessitate changes to the genome. Nine hairpin polyamides that target mouse major satellite DNA were synthesized. Their interactions with synthetic target dsDNA fragments were studied by thermal denaturation, gel-shift electrophoresis, circular dichroism, and fluorescence spectroscopy. The polyamides had different affinities for the target DNA, and fluorescent labeling of the polyamides affected their affinity for their targets. We validated the specificity of the probes in fixed cells and provide evidence that two of the probes detect target sequences in mouse living cell lines. This study demonstrates for the first time that synthetic compounds can be used for the visualization of the nuclear substructures formed by repeated DNA sequences in living cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text