Abstract.
Triple helix-forming oligonucleotides (TFOs) that specifically bind to double-stranded DNA sequences can be rationally designed, while intracellular delivery of single stranded RNA TFOs has not yet been studied in detail. In this report, we demonstrate gene and sequence-specific inhibition of MCP-1 gene expression due to interference of intracellular-generated single-stranded RNA (CU-TFO) with an overlapping SP-1/AP-1 target. Binding of synthetic 19-nucleotide (19-nt) CU-TFO to the MCP-1 promoter duplex was verified by triplex blotting. Furthermore, we confirmed binding of a 1.1-kb fusion transcript containing the 19-nt pyrimidine CU sequence to a plasmid-encoded MCP-1 promoter target duplex at pH 7.0. In tumour necrosis factor-α-stimulated HEK cells, CU-TFOs inhibited MCP-1 protein release by 76 ± 10.2% compared to intracellular-generated control oligonucleotides. Interleukin-8 as a control target gene was not affected by CU-TFO, confirming both highly specific and effective chemokine gene repression by transfectable TFO-shuttle vectors.
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Received 7 October 2004; received after revision 29 November 2004; accepted 2 December 2004
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Kautz, K., Schwarz, M. & Radeke, H.H. Interference with MCP-1 gene expression by vector generated triple helix-forming RNA oligonucleotides. CMLS, Cell. Mol. Life Sci. 62, 362–376 (2005). https://doi.org/10.1007/s00018-004-4443-y
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DOI: https://doi.org/10.1007/s00018-004-4443-y