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Construction of folate-conjugated pRNA of bacteriophage phi29 DNA packaging motor for delivery of chimeric siRNA to nasopharyngeal carcinoma cells

Gene Therapy volume 13, pages 814–820 (2006)Cite this article

A Corrigendum to this article was published on 17 October 2006

Abstract

Nasopharyngeal carcinoma is a poorly differentiated upper respiratory tract cancer that highly expresses human folate receptors (hFR). Binding of folate to hFR triggers endocytosis. The folate was conjugated into adenosine 5′-monophosphate (AMP) by 1,6-hexanediamine linkages. After reverse HPLC to reach 93% purity, the folate–AMP, which can only be used for transcription initiation but not for chain extension, was incorporated into the 5′-end of bacteriophage phi29 motor pRNA. A 16:1 ratio of folate–AMP to ATP in transcription resulted in more than 60% of the pRNA containing folate. A pRNA with a 5′-overhang is needed to enhance the accessibility of the 5′ folate for specific receptor binding. Utilizing the engineered left/right interlocking loops, polyvalent dimeric pRNA nanoparticles were constructed using RNA nanotechnology to carry folate, a detection marker, and siRNA targeting at an antiapoptosis factor. The chimeric pRNAs were processed into ds-siRNA by Dicer. Incubation of nasopharyngeal epidermal carcinoma (KB) cells with the dimer resulted in its entry into cancer cells, and the subsequent silencing of the target gene. Such a protein-free RNA nanoparticle with undetectable antigenicity has a potential for repeated long-term administration for nasopharyngeal carcinoma as the effectiveness and specificity were confirmed by ex vivo delivery in the animal trial.

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Acknowledgements

This work was supported mainly by NIH grant R01-EB03730 (Nanoscience and Nanotechnology in Biology and Medicine Program to PG), and partially by NIH grant R01-GM59944 (Institute of General Medicine to PG). We thank Taejin Lee for preparing the data of Figure 4, Philip Low for providing materials and insightful discussions, Paul Robinson for the cytometry assays, Sulma Mohammed for the animal trials, and Jeremy Hall for manuscript preparation.

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  1. Department of Pathobiology and Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA

    S Guo & P Guo

  2. Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, MS, USA

    F Huang

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Correspondence to P Guo.

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Guo, S., Huang, F. & Guo, P. Construction of folate-conjugated pRNA of bacteriophage phi29 DNA packaging motor for delivery of chimeric siRNA to nasopharyngeal carcinoma cells. Gene Ther 13, 814–820 (2006). https://doi.org/10.1038/sj.gt.3302716

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