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Enhanced specificity of HPV16 E6E7 siRNA by RNA–DNA chimera modification

Cancer Gene Therapy volume 18, pages 587–597 (2011)Cite this article

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Abstract

Although efforts have been made to develop new drugs for infectious and neoplastic diseases utilizing synthetic small interfering RNA(siRNAs), those intrinsically have undesirable effects, including silencing of unintended genes (off-target effect) and nonspecific cytotoxicity. Off-target effects can be avoided by DNA substitution in the guide strand (GS) seed region of nucleotide positions 1–8 and its complementary part of the passenger strand plus the 3′ overhang, which is designated as a double-strand RNA–DNA chimera (dsRDC). In this study, we found that the specificity of potent siRNAs targeting human papillomavirus 16 (HPV16) E6 and E7 oncogenes, which we previously reported, could be enhanced by short dsRDC modification (first six nucleotides from the 5′ end of the GS and its complementary nucleotides of the passenger strand). Such dsRDC modification reduced nonspecific cytotoxicity in two of three siRNAs (497 and 752), although not in the other (573), which correlated with their off-target effects. In addition, silencing activity was marginally impaired in two dsRDCs (497 and 573) and moderately in one (752). Finally, dsRDC-497 induced E6E7-specific growth suppression of cervical cancer cells as well as E6E7-immortalized human keratinocytes. Our results show that dsRDC modification enhances the specificity of E6E7 siRNA, which is required for use in in vivo settings.

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Acknowledgements

We thank Yukikazu Natori (Tokyo Institute of Technology) for the helpful discussion and suggestions. This work was supported in part by Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (KY) and a Grant-in-Aid for Cancer Research from the Ministry of Health Labor and Welfare of Japan (KY).

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Authors and Affiliations

  1. Section of Bacterial Pathogenesis, Graduate School of Medical and Dental Science, Tokyo Medical and Dental University, Tokyo, Japan

    K Yamato & I Nakagawa

  2. Division of Virology, National Cancer Center Research Institute Tokyo, Tokyo, Japan

    N Egawa & T Kiyono

  3. Department of Gastroenterology, Institute of Clinical Medicine, University of Tsukuba Ibaraki, Ibaraki, Japan

    S Endo & I Hyodo

  4. Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Tokyo, Japan

    K Ui-Tei & K Saigo

  5. Division of Pathology, Keio University, School of Medicine, Tokyo, Japan

    T Yamada

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  1. K Yamato
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Correspondence to K Yamato.

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Yamato, K., Egawa, N., Endo, S. et al. Enhanced specificity of HPV16 E6E7 siRNA by RNA–DNA chimera modification. Cancer Gene Ther 18, 587–597 (2011). https://doi.org/10.1038/cgt.2011.28

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  • DOI: https://doi.org/10.1038/cgt.2011.28

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