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 systemically with intravenous (I.V.) injection, and modest systemic delivery achieved with Vivo-Morpholinos by intraperitoneal (I.P.) injection. Efficient localized delivery can be achieved by injecting the Vivo-Morpholino directly into the area of interest. If you wish to initially validate the oligo in cultured cells you can use the same Vivo-Morpholino in cultures. However, unmodified Morpholinos, delivered with either electroporation or Endo-Porter, are more efficient in cell cultures than Vivo-Morpholinos.

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 uses the active component of arginine rich delivery peptides (the guanidinium group) with improved stability, low toxicity and reduced cost. The Vivo-Morpholino is assembled by coupling Vivo-Porter to a Morpholino while the oligo is still bound to its synthesis resin, allowing excellent purification by washing the solid-phase resin. Vivo-Morpholinos must be chosen prior to synthesis and cannot be added later because 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 phosphate 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 amount of oligo you will need depends on the experiment and the method of injection. For a typical 20g mouse injected at 12.5 mg/kg I.V. the dose is approximately 25 nmole per injection. This means you will get approximately 12 injections from the 300 nanomole quantity, 40 injections from the 1000 nanomole quantity and 240 injections from the 6000 nanomole quantity

The methods are simple

For best systemic delivery results the injection method of choice is I.V. although I.P. can also be used. The maximum suggested dosage in mammals is 12.5 mg/kg in a 24 hour period. This can be repeated daily. For short term experiments (3-day) Gene Tools suggests two days of I.V. injections at 12.5 mg/kg followed by analysis on day 3. 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 syringes and needles (27 to 32 gauge). 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.

You might want to compare Vivo-Morpholino dose with a bare oligo dose. The specific calculation for mg/ml concentration of your Vivo-Morpholino in terms of the oligo component (without delivery moiety) 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

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For an open-access introduction to Vivo-Morpholinos, see:

Paul A. Morcos, Yongfu Li, and Shan Jiang. Vivo-Morpholinos: A non-peptide transporter delivers Morpholinos into a wide array of mouse tissues. BioTechniques. 2008 Dec;45(6):616-26.
(access requires registration at the BioTechniques site; article access is free of charge)

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Moulton JD, Jiang S. Gene Knockdowns in Adult Animals: PPMOs and Vivo-Morpholinos. Molecules. 2009; 14(3):1304-1323.

Wu B, Li YF, Morcos PA, Doran TJ, Lu PJ and 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.

Paul A. Morcos, Yongfu Li, and Shan Jiang. Vivo-Morpholinos: A non-peptide transporter delivers Morpholinos into a wide array of mouse tissues. BioTechniques. 2008 Dec;45(6):616-26.

Li YF, Morcos PA. Design and Synthesis of Dendritic Molecular Transporter that Achieves Efficient in Vivo Delivery of Morpholino Antisense Oligo. Bioconjug Chem. 2008 Jul;19(7):1464-70. Epub 2008 Jun 20.

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 MA, USA. http://www.distrofiamuscular.net/abstracts17.htm