TY - GEN
T1 - Methylsorb
T2 - 8th International Conference on Electrical and Computer Engineering, ICECE 2014
AU - Shiddiky, Muhammad J.A.
AU - Sina, Abu Ali Ibn
AU - Carrascosa, Laura G.
AU - Palanisamy, Ramkumar
AU - Rauf, Sakandar
AU - Trau, Matt
N1 - Publisher Copyright:
© 2014 IEEE.
PY - 2015/1/28
Y1 - 2015/1/28
N2 - The analysis of DNA methylation is becoming increasingly important both in the clinic and also as a research tool to unravel key epigenetic molecular mechanisms in biology. Current methodologies for the quantification of regional DNA methylation are largely affected by comprehensive DNA sequencing methodologies which tend to be expensive, tedious, and time-consuming for many applications. Herein, we report an entirely new DNA methylation detection method referred to as 'Methylsorb', which is based on the inherent affinity of DNA bases to the gold surface (i.e., the trend of the affinity interactions is adenine > cytosine ≥ guanine > thymine) [1]. Since the degree of gold-DNA affinity interaction is highly sequence-dependent, it provides a new capability to detect DNA methylation by simply monitoring the relative adsorption of bisulfite treated DNA sequences onto a gold chip. Because the selective physical adsorption of DNA fragments to gold enable a direct read-out of regional DNA methylation, the current requirement for DNA sequencing is obviated. To demonstrate the utility of this method we present data on the regional methylation status of two CpG clusters located in the EN1 and MIR200B genes in MCF7 and MDA-MB-231 cells. The methylation status of these regions was obtained from the change in relative mass on gold surface with respect to relative adsorption of an unmethylated DNA source and this was detected using surface Plasmon resonance (SPR) in a label-free and real-time manner. We anticipate that the simplicity of this method, combined with the high level of accuracy for identifying the methylation status of cytosines in DNA, could find broad application in biology and diagnostics.
AB - The analysis of DNA methylation is becoming increasingly important both in the clinic and also as a research tool to unravel key epigenetic molecular mechanisms in biology. Current methodologies for the quantification of regional DNA methylation are largely affected by comprehensive DNA sequencing methodologies which tend to be expensive, tedious, and time-consuming for many applications. Herein, we report an entirely new DNA methylation detection method referred to as 'Methylsorb', which is based on the inherent affinity of DNA bases to the gold surface (i.e., the trend of the affinity interactions is adenine > cytosine ≥ guanine > thymine) [1]. Since the degree of gold-DNA affinity interaction is highly sequence-dependent, it provides a new capability to detect DNA methylation by simply monitoring the relative adsorption of bisulfite treated DNA sequences onto a gold chip. Because the selective physical adsorption of DNA fragments to gold enable a direct read-out of regional DNA methylation, the current requirement for DNA sequencing is obviated. To demonstrate the utility of this method we present data on the regional methylation status of two CpG clusters located in the EN1 and MIR200B genes in MCF7 and MDA-MB-231 cells. The methylation status of these regions was obtained from the change in relative mass on gold surface with respect to relative adsorption of an unmethylated DNA source and this was detected using surface Plasmon resonance (SPR) in a label-free and real-time manner. We anticipate that the simplicity of this method, combined with the high level of accuracy for identifying the methylation status of cytosines in DNA, could find broad application in biology and diagnostics.
KW - DNA adsorption
KW - DNA methylation
KW - Gold-DNA affinity
KW - Surface Plasmon resonance
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U2 - 10.1109/ICECE.2014.7027002
DO - 10.1109/ICECE.2014.7027002
M3 - Conference paper
AN - SCOPUS:84923261206
T3 - 8th International Conference on Electrical and Computer Engineering: Advancing Technology for a Better Tomorrow, ICECE 2014
SP - 17
EP - 20
BT - 8th International Conference on Electrical and Computer Engineering
PB - IEEE, Institute of Electrical and Electronics Engineers
CY - United States
Y2 - 20 December 2014 through 22 December 2014
ER -