A 5 '-Cap for DNA Probes Binding RNA Target Strands

作者：Egetenmeyer Simone; Richert Clemens^*

来源：Chemistry - A European Journal, 2011, 17(42): 11813-11827.

DOI：10.1002/chem.201101828

• DNA
• hybridization probe
• molecular caps
• molecular recognition RNA
• RNA recognition

Detecting short RNA strands with high fidelity at any of the bases of their sequence, including the termini, can be challenging, since fraying, wobbling, and refolding all compete with canonical base pairing. We performed a search for 5'-substituents of oligodeoxynucleotides that increase base pairing fidelity at the terminus of duplexes with RNA target strands. From a total of over 70 caps, differing in stacking moiety and linker, a phosphodiester-linked sequence of the residues of l-prolinol, glycine, and oxolinic acid, dubbed ogOA, was identified as a 5'-cap that stabilizes any of the four canonical base pairs, with Delta T-m values of up to + 13.1 degrees C for an octamer. At the same time, the cap increases discrimination against any of the 12 possible terminal mismatches, including mismatches that are more stable than their perfectly matched counterparts in the control duplex, such as A: A. A probe with the cap also showed increased selectivity in the detection of two closely related microRNAs, let7c and let7a, with a Delta T-m value of 9.2 degrees C. Melting curves also yielded thermodynamic data that shed light on the uniformity of molecular recognition in the sequence space of DNA: DNA and DNA: RNA duplexes. Hybridization probes with fidelity-enhancing caps should find applications in the individual and parallel detection of biologically active RNA species.

• 出版日期2011-10