Label-free and real-time sequence specific DNA detection based on supramolecular self-assembly.
A new label-free, optical method was developed to detect sequence-specific DNA based on supramolecular self-assembly. A cationic phenylene ethynylene oligomer with two pairs of positively charged side chains (OPE-2) can form a J-dimer or J-aggregate with negatively charged DNA by a combination of electrostatic and hydrophobic interactions. At microM concentrations of dsDNA (number of bases in ssDNA ranges from 8 to 32), the optimum supramolecular self-assembly occurs between OPE-2 and dsDNA and is characterized by a new absorption peak emerging at 418 nm and an increase in fluorescence intensity (about 4.5-fold for dsDNA(1)). In contrast, the self-assembled complexes between OPE-2 and ssDNA are less readily formed under the same conditions. Interestingly, the induced circular dichroism (CD) signal for OPE-2/ssDNA is quite strong, likely owing to the self-assembly onto ssDNA simultaneously templating helix formation. In contrast, the induced CD signal for OPE-2/dsDNA is weak, likely because the dsDNA is in a double helix conformation, and OPE-2 associated with the dsDNA should be outside of the helix. In fact, there is a steady decrease in the induced CD signal for ssDNA with the addition of equimolar complementary ssDNA over time that allows the monitoring of DNA hybridization in real time. Introduction of mismatched bases into the target DNA sequence prevents the full hybridization between ssDNA and the target DNA. For these cases, the decrease in the induced CD signals occurs more slowly and to a lesser extent, as some of the unhybridized portions may remain in helical association with OPE-2. In view of these observed signal changes, sequence specific DNA and single nucleotide mismatch can be detected in a very simple and sensitive manner without any modification of the DNA.
Tang
Yanli
Y
Center for Biomedical Engineering and Department of Chemical and Nuclear Engineering, University of New Mexico, Albuquerque, New Mexico 87131, USA.
Achyuthan
Komandoor E
KE
Whitten
David G
DG
D020137:Base Pair Mismatch; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D002942:Circular Dichroism; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D009693:Nucleic Acid Hybridization; D011108:Polymers / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013997:Time Factors
D004277:DNA, Single-Stranded; D011108:Polymers; D004247:DNA; C041325:polyphenylene sulfide