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Construction of a plasmid vector containing epidermal growth factor receptor and C-Jun shRNA

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Abstract

The objective of this study was to construct a plasmid vector for EGFR-hm-1 and C-Junh-825 small interfering RNA (siRNA). EGFR-hm-1 and C-Jun-hm-825 oligonucleotide fragments were synthesized and short hairpin RNA (shRNA) were amplified by PCR. Plasmids were isolated from E. coli TOP10 bacterium by restriction enzyme digestion using pst1 and BamH1 and oligonucleotide fragments were cloned into the pSilencer plasmid containing the U6 promoter. Recombinant clones were generated by transforming JM109 competent cells with plasmid vectors containing shRNA molecules. 58 base-paired EGFR-hm-1 and 59 base-paired C-Jun-hm-825 oligonucleotide fragments were isolated. The fragments were 100% homologous with human sequences available on GenBank. The recombinant pSilencer1.0 vector containing a 58-bp EGFR-hm-1 and 59-bp C-Jun-hm-825 fragment was constructed. These vectors have the potential to be used as treatment to combat skin photoaging under UV exposure.

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References

  1. Brummelkamp TR, Bernards R, Agami R (2002) A system for stable expression of short interfering RNAs in mammalian cells. Science 296:550–553. https://doi.org/10.1126/science.1068999

    Article  CAS  PubMed  Google Scholar 

  2. Gaffney DC, Soyer HP, Simpson F (2014) The epidermal growth factor receptor in squamous cell carcinoma: an emerging drug target. Australas J Dermatol 55:24–34. https://doi.org/10.1111/ajd.12025

    Article  PubMed  Google Scholar 

  3. Hwang LH, Low YC, Soares M et al (2012) Topical c-Jun silencing to prevent photoaging [abstract]. In: 29th annual meeting of the Northeastern Society of Plastic Surgeons, September 27–29, Boston. http://www.nesps.org

  4. Kim GK, Del Rosso JQ, Bellew S (2009) Skin cancer in asians: part 1: nonmelanoma skin cancer. J Clin Aesthetic Dermatol 2:39–42

    Google Scholar 

  5. Lee NS, Dohjima T, Bauer G et al (2002) Expression of small interfering RNAs targeted against HIV-1 rev transcripts in human cells. Nat Biotechnol 20:500–505. https://doi.org/10.1038/nbt0502-500

    Article  CAS  PubMed  Google Scholar 

  6. López-Camarillo C, Ocampo EA, Casamichana ML et al (2012) Protein kinases and transcription factors activation in response to UV-radiation of skin: implications for carcinogenesis. Int J Mol Sci 13:142–172. https://doi.org/10.3390/ijms13010142

    Article  PubMed  Google Scholar 

  7. Marceková Z, Flaig MJ, Kekus M et al (2010) The potential role of c-Jun activation in patients with cutaneous lichen planus. Exp Dermatol 19:74–80. https://doi.org/10.1111/j.1600-0625.2009.00965.x

    Article  PubMed  Google Scholar 

  8. Miyagishi M, Taira K (2002) U6 promoter-driven siRNAs with four uridine 3′ overhangs efficiently suppress targeted gene expression in mammalian cells. Nat Biotechnol 20:497–500. https://doi.org/10.1038/nbt0502-497

    Article  CAS  PubMed  Google Scholar 

  9. Paddison PJ, Hannon GJ (2003) siRNAs and shRNAs: skeleton keys to the human genome. Curr Opin Mol Ther 5:217–224

    CAS  PubMed  Google Scholar 

  10. Spallone G, Botti E, Costanzo A (2011) Targeted therapy in nonmelanoma skin cancers. Cancers 3:2255–2273. https://doi.org/10.3390/cancers3022255

    Article  PubMed  PubMed Central  Google Scholar 

  11. Xia H, Mao Q, Paulson HL, Davidson BL (2002) siRNA-mediated gene silencing in vitro and in vivo. Nat Biotechnol 20:1006–1010. https://doi.org/10.1038/nbt739

    Article  CAS  PubMed  Google Scholar 

  12. Zhang G, Fahmy RG, diGirolamo N, Khachigian LM (2006) JUN siRNA regulates matrix metalloproteinase-2 expression, microvascular endothelial growth and retinal neovascularisation. J Cell Sci 119:3219–3226. https://doi.org/10.1242/jcs.03059

    Article  CAS  PubMed  Google Scholar 

  13. Zhang JY, Selim MA (2012) The role of the c-Jun N-terminal Kinase signaling pathway in skin cancer. Am J Cancer Res 2:691–698

    CAS  PubMed  PubMed Central  Google Scholar 

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Correspondence to Yin Liu.

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Xiao, H., Yang, R., Yang, F. et al. Construction of a plasmid vector containing epidermal growth factor receptor and C-Jun shRNA. Arch Dermatol Res 310, 241–243 (2018). https://doi.org/10.1007/s00403-017-1803-7

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  • DOI: https://doi.org/10.1007/s00403-017-1803-7

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