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{{Biomod/2013/Aarhus/Nano_Creators/Begin|pagetype=Introduction|pagename=Introduction}}
{{Biomod/2013/Aarhus/Nano_Creators/Begin|pagetype=Introduction|pagename=Introduction}}


=='''Introduction to sisiRNA and cell penetrating peptides'''==


This part of the project deals with the conjugation of small internally segmented RNAs (sisiRNAs) to cell penetrating peptides (CPPs) and the subsequent assays of their activity. sisiRNA is a newly developed variation of the better-known siRNA which is a short, double stranded RNA molecules that is capable of inducing specific gene silencing through the RNA interference (RNAi) pathway. The [[Biomod/2013/Aarhus/Supplementary/Supplementary_Informations#Sequence_of_sisiRNA|sisiRNA design]] contains three strands, an intact guide strand and a passenger strand that consists of two shorter strands, 10 and 12 nt long. Due to the nick in the passenger strand, only the guide strand of this construct can be incorporated into the enzymatically active component of the RNAi pathway (the RISC complex), thereby greatly minimizing off-target effects. To increase the serum stability and thereby the silencing efficiency of the sisiRNA, DNA bases and locked nucleic acids (LNAs) have been inserted into the strands, thereby making it suitable for in vivo use.<cite>Bramsen</cite>
Furthermore the design includes a short strand connected to the 3’ end of the nicked passenger strand through a disulfide bond which can be used to anneal the sisiRNA to a staple strand of the DNA origami. The disulfide bond can be reduced inside the reductive cytoplasmatic environment of the cell, thereby releasing the sisiRNA from its carrier.
In order to investigate the tolerance of the sisiRNA towards heavy end modification, we performed conjugations with the polymer polyethylene glycol (PEG). This molecule is known to greatly increase the half-life of circulating siRNAs by shielding them from enzymatic attack, unspecific cell interactions and renal filtration, which would be beneficial if the construct was to be delivered without the origami.<cite>Veronese</cite>
PEG and other conjugates were linked to the nucleic acids by a reaction, referred to as a [[Biomod/2013/Aarhus/Materials_And_Methods/Methods#Click_reactions|click reaction]].


=='''Results and discussion'''==
=='''Results and discussion'''==

Revision as of 01:51, 27 May 2015

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Results and discussion

References

  1. Patel, L. N. et al. Cell penetrating peptides: Intracellular pathways and pharmaceutical perspectives. Pharm. Res. 24, 1977-92 (2007). [1]

    [Patel]

  2. Dempsey, C. E. et al. The actions of melittin on membranes. Biochim. Biophys. Acta 1031, 143-61 (1990) [1]

    [Dempsey]

  3. Li, W. et al. GALA: A designed synthetic pH-responsive amphipathic peptide with applications in drug and gene delivery. Adv. Drug Deliv. 56, 967-85 (2004) [1].

    [Li]

  4. Veronese, F. M. et al. PEGylation, successful approach to drug delivery. Drug Discov. Today '21, 1451-8 (2005) [1]

    [Veronese]

  5. Bramsen, J. B. et al. Improved silencing properties using small internally segmented interfering RNAs. Nucleic Acids Res. 35, 5886-97 (2007). [1].

    [Bramsen]

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</style> </head> <body> <div id="indexing"> <div id="sitemap"> <p id="sitemapTitle">SITEMAP | BIOMOD 2013 NANO CREATORS | Aarhus University</p> <div id="footer-contents"> <div class="footer-section"> <p class="footer-section-title">INTRODUCTION</p> <ul> <li><a href="/wiki/Biomod/2013/Aarhus">Home, abstract, animation and video</a></li> <li><a href="/wiki/Biomod/2013/Aarhus/Introduction">Introduction</a></li </ul> </div> <div class="footer-section"> <p class="footer-section-title">RESULTS AND DISCUSSION</p> <ul> <li><a href="/wiki/Biomod/2013/Aarhus/Results_And_Discussion/Origami">Origami</a></li> <li><a href="/wiki/Biomod/2013/Aarhus/Results_And_Discussion/Peptide_lock">Peptide lock</a></li> <li><a href="/wiki/Biomod/2013/Aarhus/Results_And_Discussion/Chemical_Modification">Chemical modification</a></li> <li><a href="/wiki/Biomod/2013/Aarhus/Results_And_Discussion/sisiRNA">sisiRNA</a></li> <li><a href="/wiki/Biomod/2013/Aarhus/Results_And_Discussion/System_In_Action">System in action</a></li> </ul> </div> <div class="footer-section"> <p class="footer-section-title">MATERIALS AND METHODS</p> <ul> <li><a href="/wiki/Biomod/2013/Aarhus/Materials_And_Methods/Origami">Origami</a></li> <li><a href="/wiki/Biomod/2013/Aarhus/Materials_And_Methods/Peptide_lock">Peptide lock</a></li> <li><a href="/wiki/Biomod/2013/Aarhus/Materials_And_Methods/Chemical_Modification">Chemical modification</a></li> <li><a href="/wiki/Biomod/2013/Aarhus/Materials_And_Methods/sisiRNA">sisiRNA</a></li> <li><a href="/wiki/Biomod/2013/Aarhus/Materials_And_Methods/System_In_Action">System in action</a></li> <li><a href="/wiki/Biomod/2013/Aarhus/Materials_And_Methods/Methods">Methods</a></li> </ul> </div> <div class="footer-section"> <p class="footer-section-title">SUPPLEMENTARY</p> <ul> <li><a href="/wiki/Biomod/2013/Aarhus/Supplementary/Team_And_Acknowledgments">Team and acknowledgments</a></li> <li><a href="/wiki/Biomod/2013/Aarhus/Supplementary/Optimizations">Optimizations</a></li> <li><a href="/wiki/Biomod/2013/Aarhus/Supplementary/Supplementary_Data">Supplementary data</a></li> 

                                               <li><a

href="/wiki/Biomod/2013/Aarhus/Supplementary/Supplementary_Informations">Supplementary informations</a> <li><a href="/wiki/Biomod/2013/Aarhus/Supplementary/References">References</a></li> </ul> </div> </div> <div> <p id="copyright">Copyright (C) 2013 | BIOMOD Team Nano Creators @ Aarhus University | Programming by: <a href="mailto:pvskaarup@gmail.com?Subject=BIOMOD 2013:">Peter Vium Skaarup</a>.</p> </div> </div>

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