Optical and Structural Characterization of a Chronic Myeloid Leukemia DNA Biosensor
| Autor: | João C. Lima, Dmitri I. Svergun, Milton Cordeiro, Ana Rita Otrelo-Cardoso, Pedro V. Baptista, Petr V. Konarev, Teresa Santos-Silva |
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| Rok vydání: | 2018 |
| Předmět: |
Electrophoresis
Base pair Chromosomes Human Pair 22 Fusion Proteins bcr-abl Biosensing Techniques Biochemistry Fluorescence spectroscopy chemistry.chemical_compound X-Ray Diffraction Molecular beacon Leukemia Myelogenous Chronic BCR-ABL Positive Scattering Small Angle Fluorescence Resonance Energy Transfer Humans Philadelphia Chromosome Microscale thermophoresis Chemistry Myeloid leukemia General Medicine DNA Neoplasm Förster resonance energy transfer Biophysics Molecular Medicine Nucleic Acid Conformation Chromosomes Human Pair 9 Biosensor DNA |
| Zdroj: | ACS chemical biology. 13(5) |
| ISSN: | 1554-8937 |
| Popis: | Selective base pairing is the foundation of DNA recognition. Here, we elucidate the molecular and structural details of a FRET-based two-component molecular beacon relying on steady-state fluorescence spectroscopy, small-angle X-ray scattering (SAXS), microscale thermophoresis (MST), and differential electrophoretic mobility. This molecular beacon was designed to detect the most common fusion sequences causing chronic myeloid leukemia, e14a2 and e13a2. The emission spectra indicate that the self-assembly of the different components of the biosensor occurs sequentially, triggered by the fully complementary target. We further assessed the structural alterations leading to the specific fluorescence FRET signature by SAXS, MST, and the differential electrophoretic mobility, where the size range observed is consistent with hybridization and formation of a 1:1:1 complex for the probe in the presence of the complementary target and revelator. These results highlight the importance of different techniques to explore conformational DNA changes in solution and its potential to design and characterize molecular biosensors for genetic disease diagnosis. |
| Databáze: | OpenAIRE |
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