TY - JOUR
T1 - Optical and Structural Characterization of a Chronic Myeloid Leukemia DNA Biosensor
AU - Cordeiro, Mílton
AU - Otrelo-Cardoso, Ana Rita Castro
AU - Svergun, Dmitri I.
AU - Konarev, Petr V.
AU - Lima, João Carlos
AU - Santos-Silva, Teresa
AU - Baptista, Pedro Viana
N1 - Conference code: 6th
PY - 2018/5/18
Y1 - 2018/5/18
N2 - 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.
AB - 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.
KW - X-Rays
KW - Small-angle scattering
UR - http://www.scopus.com/inward/record.url?scp=85047266335&partnerID=8YFLogxK
U2 - 10.1021/acschembio.8b00029
DO - 10.1021/acschembio.8b00029
M3 - Article
C2 - 29562136
AN - SCOPUS:85047266335
VL - 13
SP - 1235
EP - 1242
JO - ACS Chemical Biology
JF - ACS Chemical Biology
SN - 1554-8929
IS - 5
T2 - 6th Annual Conference of the International Chemical Biology Society (ICBS)
Y2 - 17 October 2017 through 20 October 2017
ER -