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TitleOptical and Structural Characterization of a Chronic Myeloid Leukemia DNA Biosensor.
Journal, issue, pagesACS Chem Biol, Vol. 13, Issue 5, Page 1235-1242, Year 2018
Publish dateMay 18, 2018
AuthorsMílton Cordeiro / Ana Rita Otrelo-Cardoso / Dmitri I Svergun / Petr V Konarev / João Carlos Lima / Teresa Santos-Silva / Pedro Viana Baptista /
PubMed AbstractSelective 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 ...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.
External linksACS Chem Biol / PubMed:29562136
MethodsSAS (X-ray synchrotron)
Structure data

SASDC95:
Light encoded DNA biosensor: e14A DNA (e14A)
Method: SAXS/SANS

SASDCA5:
Light encoded DNA biosensor: e14B DNA (e14B)
Method: SAXS/SANS

SASDCB5:
Light encoded DNA biosensor: e14C DNA (e14C)
Method: SAXS/SANS

SASDCC5:
Light encoded DNA biosensor: e14AB DNA (e14AB)
Method: SAXS/SANS

SASDCD5:
Light encoded DNA biosensor: e14ABC DNA (e14ABC)
Method: SAXS/SANS

SASDCE5:
Light encoded DNA biosensor: e13A DNA (e13A)
Method: SAXS/SANS

SASDCF5:
Light encoded DNA biosensor: e13B DNA (e13B)
Method: SAXS/SANS

SASDCG5:
Light encoded DNA biosensor: e13C DNA (e13C)
Method: SAXS/SANS

SASDCH5:
Light encoded DNA biosensor: e13AB DNA (e13AB)
Method: SAXS/SANS

SASDCJ5:
Light encoded DNA biosensor: e13ABC DNA (e13ABC)
Method: SAXS/SANS

SASDCK5:
Light encoded DNA biosensor: e13Ae14Be13C DNA (e13Ae14Be13C)
Method: SAXS/SANS

SASDCL5:
Light encoded DNA biosensor: e14Ae13Be14C DNA (e14Ae13Be14C)
Method: SAXS/SANS

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