Binary malachite green aptamer for fluorescent detection of nucleic acids.
A new probe that can fluorescently report the presence of specific nucleic acids in solution with extremely high selectivity was developed. The probe consists of malachite green-a triphenylmethane dye-and two short RNA strands, each of which comprises a fragment complementary to an analyte molecule and a fragment of a malachite green aptamer (MGA). The two RNA strands form MGA upon hybridization to the adjacent positions of the nucleic acid analyte. MGA is able to bind malachite green and enhance the fluorescence of the dye, thus monitoring the presence of the nucleic acid in solution. The probe reliably discriminates against 41 out of 42 possible single nucleotide substitutions in 14-mer DNA analyte at room temperature in physiological buffer. Consisting of unmodified RNA strands, which can be expressed in living cells, binary MGA probe represents a promising instrument for real-time nucleic acid monitoring in vivo.
Kolpashchikov
Dmitry M
DM
Division of Experimental Therapeutics, Department of Medicine, Columbia University, Box 84, 630W 168th Street, New York, New York 10032, USA. dk2111@columbia.edu
D008856:Microscopy, Fluorescence / Q000379:methods; D009696:Nucleic Acids / Q000032:analysis / Q000737:chemistry*; D012313:RNA / Q000032:analysis / Q000737:chemistry*; D012394:Rosaniline Dyes / Q000737:chemistry*; D012680:Sensitivity and Specificity
D009696:Nucleic Acids; D012394:Rosaniline Dyes; C005095:malachite green; D012313:RNA