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Zebrafish contract research and commercial licenses

For contract research using Morpholinos in zebrafish or to license Morpholinos for commercial use in zebrafish in the USA, contact Zygogen, Inc. See the Outside Resources box for more information.

Vivo-Morpholinos

Vivo-Morpholinos enter cells of adult animals


Vivo-Morpholinos take Morpholinos to the next level

Vivo-Morpholinos* are the knockdown, exon-skipping or miRNA blocking reagent of choice for in vivo experiments. Outstanding results can be achieved with intravenous (I.V.) injection, but modest results can also be obtained with Vivo-Morpholinos injected inter peritoneum (I.P.) or into localized tissue. If you wish to validate first in cultured cells, the same Vivo-Morpholino can be used in cultures.

Vivo-Morpholinos get the job done with two important ingredients

A Vivo-Morpholino is comprised of a Morpholino oligo with a unique covalently linked delivery moiety called Vivo-Porter, which is comprised of an octa-guanidine dendrimer. It encompasses the active components of arginine rich delivery peptides with low toxicity and reduced cost. The Vivo-Morpholino is assembled by coupling Vivo-Porter to a Morpholino still bound to its synthesis resin, allowing excellent purification. This also means that Vivo-Morpholinos must be chosen prior to synthesis as Vivo-Porter is added to a Morpholino prior to cleavage from its synthesis resin.

Vivo-Morpholinos are offered in three convenient amounts

All Vivo-Morpholinos are provided in 300, 1000, and 6000 nmole quantities as a liquid 0.5 mM stock (approximately 4 mg/mL dependent on sequence composition) in buffered saline ready for injection. For syntheses greater than 6000 nmol please contact Customer Support. All Vivo-Morpholinos are filtered and sterilized. For an additional fee, all Vivo-Morpholinos can be endotoxin-tested with the Charles River PTS system on site (certification included).

The methods are simple

For best results the delivery method of choice is I.V. although I.P. can also be used. The maximum suggested dosage in mammals is 15 mg/kg in a 24 hour period. This can be repeated daily. For short term experiments (4-day) Gene Tools suggests three days of injections at 12.5 mg/kg followed by analysis on day 4. Long term experiments should be optimized for your specific goals; however you can try starting with a loading dose equivalent to the short term experiment and then adjust dosage and times to find an optimum for continued delivery.

Delivery to most major organs is achieved

Following the short-term delivery experiment described above, one can expect quantifiable knockdown or exon-skipping in liver, small intestine, colon, muscle, lung, and stomach tissues with lesser but quantifiable delivery in the spleen, heart, skin and brain. For assessment of specific tissues within an organ one can achieve greater results with direct localized injections at the same concentrations described.

You don’t need much from the lab

You will need to provide your own sterile needles and syringes. You'll need 2 inch needles to reach the bottom of the vial. Vivo-Morpholinos are ready for you to inject without any other preparation.

You can perform deliveries based on the Morpholino oligo component

The molecular weight of the Morpholino oligo component is 1810 Daltons smaller than the whole Vivo-Morpholino. Your product information sheet provides the molecular weight of your Vivo-Morpholino. The 0.5 mM stock of your Vivo-Morpholino is approximately 4 mg/mL Morpholino component. A more precise calculation for mg/mL concentration of your Vivo-Morpholino in terms of the oligo component is:

mg/mL Morpholino = (Mol. Wt. Vivo-Morpholino – 1810 Da) x 0.0005

Vivo-Morpholinos are stable

Vivo-Morpholinos are provided sterile and can be stored at room temperature or put in your refrigerator. If your Vivo-Morpholino becomes contaminated, it can be autoclaved or filter sterilized. To autoclave, disable the autoclave's vacuum cycle to prevent loss of liquid. To filter sterilize, use a 0.2 micron polysulfone membrane; do not use other membranes as they have higher affinity for Vivo-Morpholinos and their use can result in significant loss of oligo concentration. To minimize decrease in oligo concentration due to association with the membrane, use the smallest available syringe filter (13 mm or smaller).

Vivo-Morpholinos can be ordered online

You can log into our online store and order Vivo-Morpholinos (to log in, see the box in the left column on this page). For more information on how to order Vivo-Morpholinos Contact Us.

*Patent pending

References

Li YF, Morcos PA. Design and Synthesis of Dendritic Molecular Transporter that Achieves Efficient in Vivo Delivery of Morpholino Antisense Oligo. Bioconjug Chem. 2008 Jun 20. [Epub ahead of print]

Wu B, Li Y, Morcos PA, Doran TJ, Lu P, Lu QL. Restoration of Dystrophin Expression in Skeletal and Cardiac Muscle by Systemic Delivery of MorpholinoE23-Vivo Porter Oligonucleotide. 2008. Abstract 433, ANNUAL MEETING OF AMERICAN SOCIETY OF GENE THERAPY, BOSTON, USA. http://www.distrofiamuscular.net/abstracts17.htm


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