RNA interference (RNAi) in plants and animals

RNA interference (RNAi) and therapeutics Name Course Instructor Date Tumor-targeted Delivery of siRNA by self-assembled Nanoparticles Introduction Accrding to Lopez-Fraga., Martinez, and Jimenez (2009), RNAi was described firstly as a molecule that regulates the mechanism of eukaryotic cells with dsRNA. After this discovery, studies in this molecule provided that RNAi can also occur in mammilla cells but by double-stranded small interfering RNAs (siRNAs) PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5Mb3Blei1GcmFnYTwvQXV0aG9yPjxZZWFyPjIwMDk8L1ll YXI+PFJlY051bT42Nzk8L1JlY051bT48RGlzcGxheVRleHQ+WzFdPC9EaXNwbGF5VGV4dD48cmVj b3JkPjxyZWMtbnVtYmVyPjY3OTwvcmVjLW51bWJlcj48Zm9yZWlnbi1rZXlzPjxrZXkgYXBwPSJF TiIgZGItaWQ9IjUwd3hkcHpkOXZkNXI3ZTl0NWI1OTVkanJmcHR0cnh3OWF2cCI+Njc5PC9rZXk+ PC9mb3JlaWduLWtleXM+PHJlZi10eXBlIG5hbWU9IkpvdXJuYWwgQXJ0aWNsZSI+MTc8L3JlZi10 eXBlPjxjb250cmlidXRvcnM+PGF1dGhvcnM+PGF1dGhvcj5Mb3Blei1GcmFnYSwgTS48L2F1dGhv cj48YXV0aG9yPk1hcnRpbmV6LCBULjwvYXV0aG9yPjxhdXRob3I+SmltZW5leiwgQS48L2F1dGhv cj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz5TeWxlbnRpcyBTQVUsIFBh cnF1ZSBUZWNub2xvZ2ljbyBkZSBNYWRyaWQsIE1hZHJpZCwgU3BhaW4uIG1sZnJhZ2EzM0BnbWFp bC5jb208L2F1dGgtYWRkcmVzcz48dGl0bGVzPjx0aXRsZT5STkEgaW50ZXJmZXJlbmNlIHRlY2hu b2xvZ2llcyBhbmQgdGhlcmFwZXV0aWNzOiBmcm9tIGJhc2ljIHJlc2VhcmNoIHRvIHByb2R1Y3Rz 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Since the discovery of siRNA, almost every gene that related to disease and has a homologous sequence has become a target for siRNA. Meany studies have been conducted in siRNA to find an effective therapy to incurable diseases such as cancers and viral infections (Lopez-Fraga, M., T. Martinez, and A. Jimenez, 2009). However, the delivery of this molecule to the target cells or tissues is the major problem that obstacle its development. Shyh, a famous researcher and his fellow team researchers have created the latest method that can be used to deliver the cells that have been attacked by cancer. Technology The self-assembled nanoparticle (NP) is the techjnology that was used to deliver siRNA by mixing with siRNA and calf thymus DNA which contains immunostimulating CpG motifs (Oh, Park, 2009). The whole complex was coated by using the polyethylene glycol-lipids so as to be able to make it more stable. The hypothesis of the delivery is protecting encapsulated siRNA by NP which then would accumulate in the lung cancer cells. In this study the lung cancer cell line (NCI-H460) was used to grow tumor in vivo (mice). For the sake of this study, the (EGFR) which is the abbreviation for Epidermal growth factor receptor. The expiration of EGER is associated with several features of tumor, such as, increasing the proliferation and decreased apoptosis. The total concentration that was needed for the NP to silence the EGFR was 120nmol/1 and there was a very positive result that was observed. The pharmacokinetic (PK) was equal in the targeted and non-targeted NP. Xenogeny IVIS imaging system was employed to display the distribution of FNM-siRNA (fluorescein-labeled siRNA) in major tissue. In order to detect, EGFR the Western blot analysis and the immunohistochemistry was applied in the experiment (Oh, Park, 2009). Application Rao (2009) argues that most of the delivery systems that developed to target cancer cells were not effective because most of injected does is reduced in the liver and lung by reticular endothelial system. For this reason this approach was developed to solve this problem. in this study, high amount of siRNA was delivered by targeted NP to silence the EGFR in NCI-H460 tumor. This result in strong gene silencing by siRNA that delivered by NP comparing with siRNA without targeted NP. On the next procedure, the blood in mice with infectious tumors eliminated the NP content firster than the blood in the mice without tumors. FAM-siRNA was delivered into intercellular more effectively by targeted NP than non targeted NP. While, the result of the distribution of siRNA by NP was 30% of fluorescence positive cells, the distribution without NP was 5%. Most of siRNA without NP remained extracellular but siRNA that delivered by NP was intercellular. Non targeted NP partly affected the tumor and only siRNA and the control did not affect the tumor. Most free siRNA did not reach the tumor because they eliminated from the blood. Tumor uptake most of NP with 70-80% ID/g while liver and lung uptake 10-20% ID/g. this is mean NPs are highly selective for tumor. In view of Rao (2009) this occurs because the size of NP is too big (120-150nm) to present in the lung and the liver but it is appropriate to insert through the capillaries in tumor tissue. siRNA did not similarly internalized in the tumor tissue because they may were taken by other angioenic capillaries cells. However, this did not affect the complete silence of EGFR. Most of immune response in the nude mice model were not because of siRNA but because CPG motifs. 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