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Screening siRNAs targeting a novel gene (HA117) and the development of a derivative recombinant adenovirus delivery system

Cancer Gene Therapy volume 18, pages 655–662 (2011)Cite this article

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Abstract

A novel gene, HA117, was discovered in our previous work. Using the pSOS-HUS vector method which we designed at previous study, we screened for small interfering RNAs (siRNAs) that targeted HA117. The pSOS-HUS siRNA screening results were verified and a delivery system was developed that contained a recombinant adenovirus carrying DNA templates for the transcription of the HA117 siRNAs. Of five pairs of DNA templates, siRNA transcribed from HAi5 produced the strongest effect against HA117. A recombinant adenovirus containing HAi5 (Ad-HAi5) was successfully constructed and evaluated. This work has laid the foundation for further study of HA117 gene function using RNA interference technology and has showed the pSOS-HUS vector method was successfully utilized as a rapid and effective screen of siRNAs for a target gene.

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References

  1. Zheng GH, Fu JR, Xu YH, Jin XQ, Liu WL, Zhou JF . Screening and cloning of multi-drug resistant genes in HL-60/MDR cells. Leuk Res 2009; 33: 1120–1123.

    Article  CAS  Google Scholar 

  2. Guo Y, Zheng G, Jin X, Xu Y, Luo Q, Liu X et al. HA117 gene increased the multidrug resistance of K562 cells in vitro: an investigation to the function of a novel gene related to drug resistance. J Exp Clin Cancer Res 2009; 28: 63.

    Article  Google Scholar 

  3. Luo Q, Kang Q, Song WX, Luu HH, Luo X, An N et al. Selection and validation of optimal siRNA target sites for RNAi-mediated gene silencing. Gene 2007; 395: 160–169.

    Article  CAS  Google Scholar 

  4. Guo Y, Jin X, Luo Q, Zheng G, Yu J, Li X et al. Construction of the recombined adenovirus vector and its expression in K562 cells. J Zhengzhou University (Med Sci) 2009; 7: 513–515.

    Google Scholar 

  5. Gopalakrishnan B, Wolff J . siRNA and DNA transfer to cultured cells. Methods Mol Biol 2009; 480: 31–52.

    Article  CAS  Google Scholar 

  6. He TC, Zhou S, da Costa LT, Yu J, Kinzler KW, Vogelstein B . A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci USA 1998; 95: 2509–2514.

    Article  CAS  Google Scholar 

  7. Sashital DG, Doudna JA . Structural insights into RNA interference. Curr Opin Struct Biol 2010; 20: 90–97.

    Article  CAS  Google Scholar 

  8. Xia H, Mao Q, Paulson HL, Davidson BL . siRNA-mediated gene silencing in vitro and in vivo. Nat Biotechnol 2002; 20: 1006–1010.

    Article  CAS  Google Scholar 

  9. Bramsen JB, Pakula MM, Hansen TB, Bus C, Langkjær N, Odadzic D et al. A screen of chemical modifications identifies position-specific modification by UNA to most potently reduce siRNA off-target effects. Nucleic Acids Res 2010; 38: 5761–5773.

    Article  CAS  Google Scholar 

  10. Strapps WR, Pickering V, Muiru GT, Rice J, Orsborn S, Polisky BA et al. The siRNA sequence and guide strand overhangs are determinants of in vivo duration of silencing. Nucleic Acids Res 2010; 38: 4788–4797.

    Article  CAS  Google Scholar 

  11. Tiemann K, Rossi JJ . RNAi-based therapeutics-current status, challenges and prospects. EMBO Mol Med 2009; 1: 142–151.

    Article  CAS  Google Scholar 

  12. Grimm D, Kay MA . Therapeutic application of RNAi: is mRNA targeting finally ready for prime time? J Clin Invest 2007; 117: 3633–3641.

    Article  CAS  Google Scholar 

  13. Rinne A, Littwitz C, Bender K, Kienitz MC, Pott L . Adenovirus-mediated delivery of short hairpin RNA (shRNA) mediates efficient gene silencing in terminally differentiated cardiac myocytes. Methods Mol Biol 2009; 515: 107–123.

    Article  CAS  Google Scholar 

  14. Kargiotis O, Chetty C, Gondi CS, Tsung AJ, Dinh DH, Gujrati M et al. Adenovirus-mediated transfer of siRNA against MMP-2 mRNA results in impaired invasion and tumor-induced angiogenesis, induces apoptosis in vitro and inhibits tumor growth in vivo in glioblastoma. Oncogene 2008; 27: 4830–4840.

    Article  CAS  Google Scholar 

  15. Chu L, Gu J, Sun L, Qian Q, Qian C, Liu X . Oncolytic adenovirus-mediated shRNA against Apollon inhibits tumor cell growth and enhances antitumor effect of 5-fluorouracil. Gene Ther 2008; 15: 484–494.

    Article  CAS  Google Scholar 

  16. Sanlioglu AD, Karacay B, Koksal IT, Griffith TS, Sanlioglu S . DcR2 (TRAIL-R4) siRNA and adenovirus delivery of TRAIL (Ad5hTRAIL) break down in vitro tumorigenic potential of prostate carcinoma cells. Cancer Gene Ther 2007; 14: 976–984.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. The Infection and Digestology Department of Affiliated Children's Hospital of Chongqing Medical University, The Children's Hospital, Chongqing, China

    G H Zheng

  2. The Surgery Department of Affiliated Children's Hospital of Chongqing Medical University, The Children's Hospital, Chongqing, China

    Q Luo & X Q Jin

  3. The Hematol Department of Affiliated Children's Hospital of Chongqing Medical University, The Pediatrics Institution of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, Key Laboratory of Pediatrics in Chongqing, CSTC2009CA5002, Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Chongqing, China

    Y X Guo & Y H Xu

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  1. G H Zheng
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  2. Q Luo
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Correspondence to Y H Xu.

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The authors declare no conflict of interest.

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Author contributions

GH Zheng drafted and revised the important intellectual content in the article; Q Luo designed and constructed the pSOS-HUS vector and directed and helped with cloning experiments; XQ Jin assisted with the conception and design of the study; YX Guo acquired, analyzed and interpreted data; YH Xu directed the concept and approved final submitted version.

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Zheng, G., Luo, Q., Jin, X. et al. Screening siRNAs targeting a novel gene (HA117) and the development of a derivative recombinant adenovirus delivery system. Cancer Gene Ther 18, 655–662 (2011). https://doi.org/10.1038/cgt.2011.32

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  • DOI: https://doi.org/10.1038/cgt.2011.32

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