SASDCB5: Light encoded DNA biosensor: e14C DNA (e14C)

Basic information

• Entry: Database: SASBDB / ID: SASDCB5

Sample

Light encoded DNA biosensor: e14C DNA

• e14C (DNA)

• Citation: Journal: ACS Chem Biol / Year: 2018; Title: Optical and Structural Characterization of a Chronic Myeloid Leukemia DNA Biosensor.; Authors: Mílton Cordeiro / Ana Rita Otrelo-Cardoso / Dmitri I Svergun / Petr V Konarev / João Carlos Lima / Teresa Santos-Silva / Pedro Viana Baptista / ; Abstract: 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.

Contact author

• Petr Konarev (EMBL-Hamburg, European Molecular Biology Laboratory (EMBL) - Hamburg outstation, Notkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany)

Models

• Model #1331: ; Type: dummy / Radius of dummy atoms: 1.00 A / Chi-square value: 0.906 / P-value: 0.391510; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: Light encoded DNA biosensor: e14C DNA / Specimen concentration: 1 mg/ml
• Buffer: Name: 154 mM NaCl / pH: 8.3

Entity #711

Type: DNA / Description: e14C / Formula weight: 5.496 / Num. of mol.: 1
Sequence:

ccacgccaaa ccacgaaa

Experimental information

• Beam: Instrument name: ESRF BM29 / City: Grenoble / 国: France / Type of source: X-ray synchrotronSynchrotron / Wavelength: 0.1 Å / Dist. spec. to detc.: 3 mm
• Detector: Name: Pilatus 1M

Scan

Title: Light encoded DNA biosensor: e14C DNA / Measurement date: Jun 16, 2016 / Storage temperature: 10 °C / Cell temperature: 10 °C / Exposure time: 0.05 sec. / Number of frames: 20 / Unit: 1/nm /

	Min	Max
Q	0.0771	4.9518

Distance distribution function P(R)

Sofotware P(R): GNOM 4.6 / Number of points: 512 /

	Min	Max
Q	0.1266	4.949
P(R) point	20	531
R	0	5

Result

Experimental MW: 6 kDa / Type of curve: merged

	P(R)	Guinier	Guinier error
Forward scattering, I0	10.71	10.65	0.016
Radius of gyration, Rg	1.29 nm	1.25 nm	0.05

	Min	Max
D	-	5
Guinier point	16	203