14.06.2012

Meeting with Dr. Zhang and discussion about the project People: Alexander, Karen, Svea, Praveen, Ali, Santiago, Varsha, Agata, Thomas, Dr. Zhang Time: 17:30

1. Alexander presented the Steps for the experiment-> Find presentation in the googlegroup
   1. We need three different cholesterol labelled oligos
   2. 9 connections to anchor the Origami on the outside of the GUV
   3. Ask Dominik to use the origami structure anchored to a guv (already in press in the paper);
   4. B CUBE has a pool of 30 different staples/oligos coupled to cholesterol;
   5. One cholesterol is added to one oligonucleotide: the big challenge is the solubility, with more cholesterols added
   6. There is no good method to purify it because the cholesterols would cluster together and the oligos stay outside, the cholesterol is added at the 3’ end; so the staples hanging out must be also complementary at the 3’ end.
   7. Origami coupling to the GUV: combining one anchor strand with a staple

*Discussion about getting rid of one of the lids, so we would not have the problem with a possible sterical hindrance, could be possible with different lock systems at the correct/proper places *How many origamis? Coat the GUV with LUVs, so nearly coat the origami completely *How much time is needed to synthetize the cholesterol coupled oligos? *In B-cube (Dr. Zhang) they have already a lot of coupled oligos (in storage) and for the coupling you really need not many… *Problem with connecting GUVs for the control (because no cholesterol labelled complementary strands): oligos can form hairpin structures so it should be possible; have a linker oligo *We have to think about a partly double stranded structure to stabilize it *Meeting tomorrow at noon: at B-cube (to discuss with Svea and Michael Schlierf about the control experiment and the attachment of the two GUVs ) *Karen will ask Dominik Kauert if he has any oligos left (stock solutions) which are labelled with cholesterol and which one for the sequence