Vivo-Morpholinos for adult animals
Jon Moulton
Tuesday, 12 February 2008 00:49 UTC
The availability of Vivo-Morpholinos makes the specific and effective activities of Morpholino oligos available in many animal model systems where Morpholinos were not effective in the past due to delivery issues. Highly specific Morpholino translation blocking, splice blocking and miRNA maturation inhibition are now feasible in adult rodents, allowing knockdowns without the known widespread off-target gene modulation associated with RNAi technologies and RNase-H dependent antisense.
Vivo-Morpholinos enter the cells of adult organisms when administered by I.V. injection. Initial results in mouse, rat and adult zebrafish show activity of splice-blocking and translation-blocking Morpholinos in tissues after introduction of Vivo-Morpholinos into the blood. Delivery efficacy varies by tissue type, but this is far better activity than is available from bare Morpholino oligos.
Vivo-Morpholinos are Morpholino oligos covalently linked to octaguanidinium dendrimers. The dendrimers are synthesized on the Morpholino oligo while the oligos are still attached to their synthesis resin, then capped with guanidiniums. The oligos are then cleaved from the resin and the bases deprotected. This means that existing oligos cannot be modified to be Vivo-Morpholinos; addition of a delivery moiety must be specified prior to synthesis. Vivo-Morpholinos are available on the Gene Tools online store and are described on the Gene Tools website (http://www.gene-tools.com/vivomorpholinos), where you can find links to some gel images from splice-modification experiments in mouse.
After many years of research, the folks at Gene Tools are very excited to finally offer a delivery moiety that brings Morpholino techniques to life science researchers working in adult animals.
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Replies
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Hi Jon,
Sounds good – I looked at the GeneTools homepage but did not see any data or references for zebrafish. You mention the use of the in vivo morpholinoes in zebrafish – could you tell me how these were applied to the fish (in the blood?) and how it worked in this model. I could imagine to use it as miRNA blocker or as an alternative to siRNAs – but I guess they are expensive?
Brian -
Hi Brian,
There’s no publication yet and our only data are some images from a collaborator, which we can’t post since he might publish. The oligos were indeed injected into the blood of the fish, and had the expected phenotypic effect.
For prices, combine custom Morpholino with a 3’-Vivo-Porter: http://www.gene-tools.com/node/15
They are considerably more expensive than a bare oligo, but less than double the price. We do a multi-step synthesis on the oligo while its still bound to its synthesis resin, then cleave the modified oligo from the resin while deprotecting the bases, then go through some additional purification steps and dissolve the oligo in physiological saline. Vivo-Porter is shipped sterile in an ampule with a pierceable septum.
The first publication is still a few months off. When it appears, I’ll post the citation here in the Morpholino group.
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Great – thanks for all the info. We will consider it!
Brian -
The publications are starting to appear.
Wu B, Li Y, Morcos PA, Doran TJ, Lu P, Lu QL. Octa-guanidine Morpholino Restores Dystrophin Expression in Cardiac and Skeletal Muscles and Ameliorates Pathology in Dystrophic mdx Mice. Mol Ther. 2009 May;17(5):864-71. Epub 2009 Mar 10.
http://www.nature.com/mt/journal/v17/n5/full/mt200938a.html
(full text now onilne, open access)
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