Chemical Synthesis of LNA-mCTP and its application for MicroRNA detection

Nucleosides, Nucleotides & Nucleic Acids
Anilkumar R KoreZhongting Hu

Abstract

Locked nucleic acids (LNA) are being applied in hybridization studies, but current locked nucleotides cannot be transcribed into RNA probes. Here, the authors report the use of a new synthetic locked nucleotide, locMeCytidine-5'-triphosphate (LNA-mCTP), for hybridization study. This synthetic LNA-mCTP can be transcribed into a short ( approximately 30-nt) RNA probe. Dot blot hybridization on nylon membrane suggested that the short (33)P-LNA RNA probes had strong binding affinity to target oligonucleotides and its detection sensitivity was approximately approximately 1000 miRNAs in a 20- to 30-mum (diameter) dot area. On tissue sections, the differential expression pattern of mir-124 within different tissue regions revealed by short (33)P- LNA RNA probes correlated well to that analyzed by real-time RT-PCR. In addition, the specific cellular distribution of vasoactive intestinal polypeptide mRNAs in the mouse brain was the same using a 30-nt (33)P-LNA RNA probe and a 1.5-kb (33)P-RNA probe. These results suggested the high hybridization specificity of the small LNA-RNA probes to target small RNAs. Finally, the authors applied (33)P-LNA probes to detect miRNA let-7C expression in human cancer tissues. Let-7C was clearly present i...Continue Reading

Citations

Mar 21, 2009·Organic & Biomolecular Chemistry·Rakesh N VeeduJesper Wengel
Jul 16, 2008·Journal of Cellular and Molecular Medicine·Xu-Bao ShiRalph W Devere White
Dec 14, 2011·Chembiochem : a European Journal of Chemical Biology·Lasse H LauridsenRakesh N Veedu
May 9, 2012·Wiley Interdisciplinary Reviews. RNA·Pauline Chugh, Dirk P Dittmer
Jun 7, 2008·Journal of the American Chemical Society·Rakesh N VeeduJesper Wengel
Oct 27, 2021·Nucleosides, Nucleotides & Nucleic Acids·Muthian ShanmugasundaramAnilkumar R Kore

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