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Induction of Transcriptional Gene Silencing by Expression of shRNA Directed to c-Myc P2 Promoter in Hepatocellular Carcinoma by Tissue-Specific Virosomal Delivery

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Promoter Associated RNA

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1543))

Abstract

Double-stranded RNA-mediated transcriptional gene silencing (TGS) has shown promising results over posttranscriptional gene silencing (PTGS) due to its long term and heritable nature. Various research groups have shed light on different mechanisms by which TGS operate. Some of these include histone modification, DNA methylation, or restriction of RNA polymerase binding onto the target gene’s promoter. This serves as an added advantage since permanent c-Myc inactivation is critical for suppressing hepatocellular carcinoma (HCC). Inability to target cancer cells specifically, without affecting the normal cells, has been one of the biggest drawbacks of an effective cancer therapy. Therefore, we aimed to overcome this barrier by first generating tumor-specific transcriptional units expressing TGS inducing shRNAs against c-Myc’s P2 promoter only in neoplastic liver cells. Secondly, we coupled this TGS inducing system with Sendai fusion virosomes for liver-specific delivery to minimize nonspecific side effects in vitro.

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Acknowledgment

This work was supported by a grant from Department of Biotechnology, Government of India (Grant No BT/PR13733/AGR/36/667/2010) and BMC Cancer (Biomedcentral) for permitting the reuse of already published data by MKZ (DOI:10.1186/1471-2407-14-582). Department of Science and Technology’s J.C. Bose fellowship and R&D grant from Delhi University to Professor Debi P. Sarkar is also acknowledged. Mohammad Khalid Zakaria was supported by a research fellowship from Indian Council of Medical Research (I.C.M.R).

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Authors and Affiliations

  1. Department of Biochemistry, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029, India

    Mohammad Khalid Zakaria & Parthaprasad Chattopadhyay

  2. International Centre for Genetic Engineering and Biotechnology, Padriciano 99, Trieste, 34149, Italy

    Mohammad Khalid Zakaria

  3. Department of Biochemistry, University of Delhi, South Campus, New Delhi, 110021, India

    Debi P. Sarkar

Authors
  1. Mohammad Khalid Zakaria
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  2. Debi P. Sarkar
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  3. Parthaprasad Chattopadhyay
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Corresponding author

Correspondence to Mohammad Khalid Zakaria .

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Editors and Affiliations

  1. Experimental Therapeutics Group, Institute of Oncology Research, Bellinzona, Switzerland

    Sara Napoli

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Zakaria, M.K., Sarkar, D.P., Chattopadhyay, P. (2017). Induction of Transcriptional Gene Silencing by Expression of shRNA Directed to c-Myc P2 Promoter in Hepatocellular Carcinoma by Tissue-Specific Virosomal Delivery. In: Napoli, S. (eds) Promoter Associated RNA. Methods in Molecular Biology, vol 1543. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6716-2_14

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  • DOI: https://doi.org/10.1007/978-1-4939-6716-2_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6714-8

  • Online ISBN: 978-1-4939-6716-2

  • eBook Packages: Springer Protocols

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