Citations of papers using Vivo-Morpholinos are here.
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, which is comprised of an octa-guanidine dendrimer. It uses the active component of arginine rich delivery peptides (the guanidinium group) with improved stability and reduced cost. The Vivo-Morpholino is assembled by coupling the vivo-delivery group 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 the vivo-delivery group 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 400, 2000, and 10000 nmole quantities as a liquid 0.5 mM stock in phosphate buffered saline ready for injection. For syntheses greater than 10000 nmol please contact Customer Support. All Vivo-Morpholinos are filtered and sterilized. 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 16 injections from the 400 nanomole quantity, 80 injections from the 2000 nanomole quantity and 400 injections from the 10000 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; dosing can be repeated daily, however, the sensitivity of organisms to the toxicity of the delivery moiety can vary depending on the type and health of the organism, so it is important to check the tolerance of your organism prior to dosing a group of animals.
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. Vivo-Morpholinos were optimized using 4 to 10 week old C57 Black mice and up to 20 mg/kg injections with successful outcomes. However, we have found that younger or older mice do not tolerate Vivo-Morpholinos as well and may require substantially lower doses. In addition the use of compromised mice, such as those with less robust genetic backgrounds, may require further dose limitations.
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.
*US Patent 7,935,816
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.
Renzi A, Mancinelli R, Onori P, Franchitto A, Alpini G, Glaser S, Gaudio E. Inhibition of the liver expression of arylalkylamine N-acetyltransferase increases the expression of angiogenic factors in cholangiocytes. Hepatobiliary Surg Nutr. 2014;3(1):4-10. doi:10.3978/j.issn.2304-3881.2014.01.02
Shaked I, Hanna DB, Gleißner C, Marsh B, Plants J, Tracy D, Anastos K, Cohen M, Golub ET, Karim R, Lazar J, Prasad V, Tien PC, Young MA, Landay AL, Kaplan RC, Ley K. Macrophage Inflammatory Markers Are Associated With Subclinical Carotid Artery Disease in Women With Human Immunodeficiency Virus or Hepatitis C Virus Infection. Arterioscler Thromb Vasc Biol. 2014 Mar 20. [Epub ahead of print]
Nizzardo N, Simone C, Salani S, Ruepp M-D, Rizzo F, Ruggieri M, Zanetta C, Brajkovic S, Moulton HM, Müehlemann O, Bresolin N, Comi GP, Corti S. Effect of Combined Systemic and Local Morpholino Treatment on the Spinal Muscular Atrophy Δ7 Mouse Model Phenotype. Clin Therap. 2014. 36(3):340-56. doi:10.1016/j.clinthera.2014.02.004
Carraro G, Shrestha A, Rostkovius J, Contreras A, Chao CM, El Agha E, Mackenzie B, Dilai S, Guidolin D, Taketo MM, Günther A, Kumar ME, Seeger W, De Langhe S, Barreto G, Bellusci S. miR-142-3p balances proliferation and differentiation of mesenchymal cells during lung development. Development. 2014 Feb 19. [Epub ahead of print]
Glaser S, Meng F, Han Y, Onori P, Chow BK, Francis H, Venter J, McDaniel K, Marzioni M, Invernizzi P, Ueno Y, Lai JM, Huang L, Standeford H, Alvaro D, Gaudio E, Franchitto A, Alpini G. Secretin Stimulates Biliary Cell Proliferation by Regulating Expression of MicroRNA 125b and MicroRNA let7a in Mice. Gastroenterology. 2014 Feb 25. pii: S0016-5085(14)00241-8. doi: 10.1053/j.gastro.2014.02.030. [Epub ahead of print]
Two tail-vein injections at 30 mg/kg targeting microRNAs mmu-miR-125b or mmu-miR-let7
Reissner KJ, Gipson CD, Tran PK, Knackstedt LA, Scofield MD, Kalivas PW. Glutamate transporter GLT-1 mediates N-acetylcysteine inhibition of cocaine reinstatement. Addic Biol. 2014;[Epub ahead of print] doi:10.1111/adb.12127
Rat nucleus accumbens
Lee TKW, Cheung VCH, Lu P, Lau EYT, Ma S, Tang KH, Tong M, Lo J, Ng IOL. Blockade of CD47 mediated CTSS-PAR2 signaling provides a therapeutic target for hepatocellular carcinoma. Hepatology. 2014;[Epub ahead of print] doi:10.1002/hep.27070
Human xenograft in mice, intratumoral injection
Ferguson DP, Dangott LJ, Schmitt EE, Vellers HL, Lightfoot JT. Differential Skeletal Muscle Proteome of High and Low Active mice. J Appl Physiol. 1985. 2014 Feb 6. [Epub ahead of print]
Karkampouna S, Kruithof BP, Kloen P, Obdeijn MC, van der Laan AM, Tanke HJ, Kemaladewi DU, Hoogaars WM, 't Hoen PA, Aartsma-Rus A, Clark IM, Ten Dijke P, Goumans MJ, Kruithof-de Julio M. Novel Ex Vivo Culture Method for the Study of Dupuytren's Disease: Effects of TGFβ Type 1 Receptor Modulation by Antisense Oligonucleotides. Mol Ther Nucleic Acids. 2014 Jan 21;3:e142. doi: 10.1038/mtna.2013.69.
Human explant cultures
Velu CS, Chaubey A, Phelan JD, Horman SR, Wunderlich M, Guzman ML, Jegga AG, Zeleznik-Le NJ, Chen J, Mulloy JC, Cancelas JA, Jordan CT, Aronow BJ, Marcucci G, Bhat B, Gebelein B, Grimes HL. Therapeutic antagonists of microRNAs deplete leukemia-initiating cell activity. J Clin Invest. 2014 Jan 2;124(1):222-36. doi: 10.1172/JCI66005. Epub 2013 Dec 16.
Bollinger RJ, Bujak, JK, Madsen SS, Tipsmark CK. Vivo-Morpholino Induced Knock-Down of Gill Na,K-ATPase Impairs Seawater Acclimation in Rainbow Trout. Society for Integrative and Comparative Biology 2014. (poster) P2.136 firstname.lastname@example.org
Khandekar G, Jagadeeswaran P. Role of hepsin in factor VII activation in zebrafish. Blood Cells Mol Dis. 2013 Aug 15. doi:pii: S1079-9796(13)00194-0. 10.1016/j.bcmd.2013.07.014. [Epub ahead of print]
Bitar MS, Abdel-Halim SM, Al-Mulla F. Caveolin-1 upregulation in diabetic fibroblasts and wounded tissues: implication for understanding the underlying mechanisms of non-healing diabetic ulcers. Am J Physiol Endocrinol Metab. 2013 Aug 13. [Epub ahead of print]
Gao Z, Cooper TA. Reexpression of pyruvate kinase M2 in type 1 myofibers correlates with altered glucose metabolism in myotonic dystrophy. PNAS USA. 2013; [Epub ahead of print] doi:10.1073/pnas.1308806110
Voskoboynik A, Newman AM, Corey DM, Sahoo D, Pushkarev D, Neff NF, Passarelli B, Koh W, Ishizuka KJ, Palmeri KJ, Dimov IK, Keasar C, Fan HC, Mantalas GL, Sinha R, Penland L, Quake SR, Weissman IL. Identification of a colonial chordate histocompatibility gene. Science. 2013 Jul 26;341(6144):384-7. doi: 10.1126/science.1238036.
Knockdown in Botryllus schlosseri,a colonial urochordate
Spatazza J, Lee HH, Di Nardo AA, Tibaldi L, Joliot A, Hensch TK, Prochiantz A. Choroid-Plexus-Derived Otx2 Homeoprotein Constrains Adult Cortical Plasticity. Cell Rep. 2013 Jun 11. doi:pii: S2211-1247(13)00231-3. 10.1016/j.celrep.2013.05.014. [Epub ahead of print]
Kizil C, Iltzsche A, Kaslin J, Brand M. Micromanipulation of Gene Expression in the Adult Zebrafish Brain Using Cerebroventricular Microinjection of Morpholino Oligonucleotides. J Vis Exp. 2013;(75):e50415, doi:10.3791/50415
Video protocol for cerebroventricular microinjection in adult zebrafish.
Martin NT, Nakamura K, Davies R, Nahas SA, Brown C, Tunuguntla R, Gatti RA, Hu H. ATM-Dependent MiR-335 Targets CtIP and Modulates the DNA Damage Response. PLoS Genet. 2013;9(5):e1003505. doi:10.1371/journal.pgen.1003505
Lymphoblastoid cell lines
Jacobi CLJ, Rudigier LJ, Scholz H, Kirschner KM. Transcriptional regulation by the Wilms tumor protein, Wt1, suggests a role of the metalloproteinase Adamts16 in murine genitourinary development. J Biol Chem. 2013;[Epub ahead of print] doi:10.1074/jbc.M113.464644
Murine organotypic cultures: kidney and gonads
Ferguson DP, Schmitt EE, Lightfoot JT. Vivo-Morpholinos Induced Transient Knockdown of Physical Activity Related Proteins. PLoS ONE. 2013;8(4):e61472. doi:10.1371/journal.pone.0061472
Mouse; bradykinin enhancing brain delivery
Stec DE, Storm MV, Pruett BE, Gousset MU. Antihypertensive Actions of Moderate Hyperbilirubinemia: Role of Superoxide Inhibition. Am J Hypertens. 2013 Mar 12. [Epub ahead of print] doi: 10.1093/ajh/hpt038
Kizil C, Kyritsis N, Dudczig S, Kroehne V, Freudenreich D, Kaslin J, Brand M. Regenerative neurogenesis from neural progenitor cells requires injury-induced expression of Gata3. Dev Cell. 2012. 23(6):1230-7. doi: 10.1016/j.devcel.2012.10.014.
Kyritsis N, Kizil C, Zocher S, Kroehne V, Kaslin J, Freudenreich D, Iltzsche A, Brand M. Acute inflammation initiates the regenerative response in the adult zebrafish brain. Science. 2012 Dec 7;338(6112):1353-6. doi: 10.1126/science.1228773.
Lin JF, Pan HC, Ma LP, Shen YQ, Schachner M. The Cell Neural Adhesion Molecule Contactin-2 (TAG-1) Is Beneficial for Functional Recovery after Spinal Cord Injury in Adult Zebrafish. PLoS ONE. 2012;7(12):e52376. doi:10.1371/journal.pone.0052376 Vivo-Morpholino in Gelfoam at spinal cord injury: "Contactin-2 anti-sense MO significantly reduced contactin-2 protein levels by approximately 82% compared with control MO 10 days after MO treatment being set to 100% ... . Even 3 weeks after MO treatment, contactin-2 protein levels continued to remain decreased to a level of 49% (data not shown). It is noteworthy in this context that MO inhibition has been observed to last for more than 6 weeks."
Hughes CE, Radhakrishnan UP, Lordkipanidzé M, Egginton S, Dijkstra JM, Jagadeeswaran P, Watson SP. G6f-Like Is an ITAM-Containing Collagen Receptor in Thrombocytes. PLoS ONE. 2012;7(12):e52622. doi:10.1371/journal.pone.0052622
Vivo-Morpholinos injected into both adult zebrafish (>3 month) and zebrafish embryos (3dpf).
Malerba A, Kang JK, McClorey G, Saleh AF, Popplewell L, Gait MJ, Wood MJA, Dickson G. Dual Myostatin and Dystrophin Exon Skipping by Morpholino Nucleic Acid Oligomers Conjugated to a Cell-penetrating Peptide Is a Promising Therapeutic Strategy for the Treatment of Duchenne Muscular Dystrophy. Mol Ther Nucleic Acids. 2012;1:e62. doi:/10.1038/mtna.2012.54
Frampton G, Ueno Y, Quinn M, McMillin M, Pae HY, Galindo C, Leyva-Illades D, Demorrow S. The novel growth factor, progranulin, stimulates mouse cholangiocyte proliferation via Sirtuin1-mediated inactivation of FOXO1. Am J Physiol Gastrointest Liver Physiol. 2012 Oct 18. [Epub ahead of print]
Vivo-Morpholino injected into mice.
Renzi A, Demorrow S, Onori P, Carpino G, Mancinelli R, Meng F, Venter J, White M, Franchitto A, Francis H, Han Y, Ueno Y, Dusio G, Jensen KJ, Greene JJ, Glaser S, Gaudio E, Alpini G. Modulation of the biliary expression of arylalkylamine N-acetyltransferase alters the autocrine proliferative responses of cholangiocytes. Hepatology. 2012 Oct 18. doi: 10.1002/hep.26105. [Epub ahead of print]
Vivo-Morpholino injected into rats.
Csányi G, Yao M, Rodriguez AI, Al Ghouleh I, Sharifi-Sanjani M, Frazziano G, Xiaojun H, Kelley EE, Isenberg JS, Pagano PJ. Thrombospondin-1 Regulates Blood Flow via CD47 Receptor–Mediated Activation of NADPH Oxidase 1. Arterioscler Thromb Vasc Biol. 2012;[Epub ahead of print] doi:10.1161/ATVBAHA.112.300031
Luo YJ, Su YH. Opposing Nodal and BMP Signals Regulate Left–Right Asymmetry in the Sea Urchin Larva. PLoS Biol. 2012;10(10): e1001402. doi:10.1371/journal.pbio.1001402
Chen SX, Cherry A, Tari PK, Podgorski K, Kwong YKK, Haas K. The Transcription Factor MEF2 Directs Developmental Visually Driven Functional and Structural Metaplasticity. Cell. 2012;151(1):41–55 doi:10.1016/j.cell.2012.08.028
Domínguez-Bendala J, Álvarez-Cubela S, Nieto M, Vargas N, Espino-Grosso P, Sacher VY, Pileggi A, García E, Ricordi C, Inverardi L, Pastori RL. Intracardial Embryonic Delivery of Developmental Modifiers In Utero. Cold Spring Harb Protoc. 2012;doi:10.1101/pdb.prot069427
Vivo-Morpholino targeting miRNA in mouse embryos.
Xu H, Oliveira-Sales EB, McBride F, Liu B, Hewinson J, Toward M, Hendy EB, Graham D, Dominiczak AF, Giannotta M, Waki H, Ascione R, Paton JF, Kasparov S. Upregulation of junctional adhesion molecule-A is a putative prognostic marker of hypertension. Cardiovasc Res. 2012 Aug 22. [Epub ahead of print]
Tail vein injection in juvenile rats.
Chen Y, Londraville R, Brickner S, El-Shaar L, Fankhauser K, Dearth C, Fulton L, Sochacka A, Bhattarai S, Marrs JA, Liu Q. Protocadherin-17 function in zebrafish retinal development. Dev Neurobiol. 2012 Aug 28. doi: 10.1002/dneu.22053. [Epub ahead of print]
Injection into eyes of zebrafish embryos.
Pal D, Dasgupta S, Kundu R, Maitra S, Das G, Mukhopadhyay S, Ray S, Majumdar SS, Bhattacharya S. Fetuin-A acts as an endogenous ligand of TLR4 to promote lipid-induced insulin resistance. Nat Med. 2012 Aug;18(8):1279-85. doi: 10.1038/nm.2851. Epub 2012 Jul 29
Vivo-MO to FetA or Tlr4, 5- to 6-week-old female BALB/c mice, "Twenty-five nmoles of VMO was delivered to tail vein through injection in each mouse for 5 consecutive days, one on each day."
Yokota T, Nakamura A, Nagata T, Saito T, Kobayashi M, Aoki Y, Echigoya Y, Partridge T, Hoffman EP, Takeda S. Extensive and Prolonged Restoration of Dystrophin Expression with Vivo-Morpholino-Mediated Multiple Exon Skipping in Dystrophic Dogs. Nucleic Acid Therapeutics. 2012. [Epub ahead of print] doi:10.1089/nat.2012.0368
Lai BS, Witola WH, El Bissati KE, Zhou Y, Mui E, Fomovska A, McLeod R. Molecular target validation, antimicrobial delivery, and potential treatment of Toxoplasma gondii infections. PNAS. 2012;[Epub ahead of print] doi:10.1073/pnas.1208775109
Aoki Y, Yokota T, Nagata T, Nakamura A, Tanihata J, Saito T, Duguez SMR, Nagaraju K, Hoffman EP, Partridge T, Takeda S. Bodywide skipping of exons 45–55 in dystrophic mdx52 mice by systemic antisense delivery. PNAS. 2012;[Epub ahead of print] doi:10.1073/pnas.1204638109
Cutler CP, Smith D. Vivo-morpholino knockdown of aquaporin 1 (AQP1) protein expression in American eel (Anguilla rostrata) gastrointestinal tract. The Bulletin, MDI Biological Laboratory. 2012;51
Notch EG, Chapline C, Flynn E, Lameyer T, Lowell A, Sato D, Shaw JR, Stanton BA. Mitogen activated protein kinase 14-1 regulates serum glucocorticoid kinase 1 during seawater acclimation in Atlantic killifish, Fundulus heteroclitus. Comp Biochem Physiol A Mol Integr Physiol. 2012 May 2. [Epub ahead of print]
Nazmi A, Mukhopadhyay R, Dutta K, Basu A. STING Mediates Neuronal Innate Immune Response Following Japanese Encephalitis Virus Infection. Sci Rep. 2012;2:347. Epub 2012 Apr 2.
3–4 weeks old BALB/c mice, 5 mg/kg Vivo-Morpholino once per day for five days.
Parra MK, Gallagher TL, Amacher SL, Mohandas N, Conboy JG. Deep intron elements mediate nested splicing events at consecutive AG-dinucleotides to regulate alternative 3' splice site choice in vertebrate 4.1 genes. Mol Cell Biol. 2012 Apr 2. [Epub ahead of print]
Vivo-Morpholinos injected in mouse tail veins at 15mg/kg on two consecutive days.
Sartor GC, Aston-Jones GS. A Septal-Hypothalamic Pathway Drives Orexin Neurons, Which Is Necessary for Conditioned Cocaine Preference. J Neurosci. 2012;32(13):4623-31. doi:10.1523/JNEUROSCI.4561-11.2012
Vivo-Morpholinos administered though cannulae into rat brains.
Owen LA, Uehara H, Cahoon J, Huang W, Simonis J, Ambati BK. Morpholino-Mediated Increase in Soluble Flt-1 Expression Results in Decreased Ocular and Tumor Neovascularization. PLoS ONE. 2012;7(3):e33576. doi:10.1371/journal.pone.0033576
Liu Y, Yu H, Deaton SK, Szaro BG. Heterogeneous Nuclear Ribonucleoprotein K, an RNA-Binding Protein, Is Required for Optic Axon Regeneration in Xenopus laevis. J Neurosci. 2012;32(10):3563-3574. doi:10.1523/JNEUROSCI.5197-11.2012
"In uninjured eyes, it efficiently knocked down hnRNP K expression in only the RGCs, without inducing either an axotomy response or axon degeneration."
Jakobson M, Lintulahti A, Arumäe U. mRNA for N-Bak, a neuron-specific BH3-only splice isoform of Bak, escapes nonsense-mediated decay and is translationally repressed in the neurons. Cell Death Dis. 2012 Feb 2;3:e269. doi: 10.1038/cddis.2012.4.
Cultured mouse neurons
Nakamura K, Du L, Tunuguntla R, Fike F, Cavalieri S, Morio T, Mizutani S, Brusco A, Gatti RA. Functional characterization and targeted correction of ATM mutations identified in Japanese patients with ataxia-telangiectasia. Hum Mutat. 2012 Jan;33(1):198-208. doi: 10.1002/humu.21632. Epub 2011 Nov 9.
Cultured primary cells
Notch EG, Shaw JR, Coutermarsh BA, Dzioba M, Stanton BA. Morpholino Gene Knockdown in Adult Fundulus heteroclitus: Role of SGK1 in Seawater Acclimation. PLoS ONE. 2011;6(12): e29462. doi:10.1371/journal.pone.0029462
Intraperitoneal (IP) injection, 14 µg/g SGK1 translation-blocking vivo-morpholino.
Taniguchi-Ikeda M, Kobayashi K, Kanagawa M, Yu CC, Mori K, Oda T, Kuga A, Kurahashi H, Akman HO, DiMauro S, Kaji R, Yokota T, Takeda S, Toda T. Pathogenic exon-trapping by SVA retrotransposon and rescue in Fukuyama muscular dystrophy. Nature. 2011 Oct 5;478(7367):127-31. doi: 10.1038/nature10456.
Tail-vein and local injections in a mouse model of Fukuyama muscular dystrophy
Kizil C, Brand M. Cerebroventricular Microinjection (CVMI) into Adult Zebrafish Brain Is an Efficient Misexpression Method for Forebrain Ventricular Cells. PLoS ONE. 2011;6(11):e27395. doi:10.1371/journal.pone.0027395
Injection of up to 500 nl into cerebroventricular fluid at Vivo-Morpholino concentrations up to 500 µM.
Osorio FG, Navarro CL, Cadiñanos J, López-Mejía IC, Quirós PM, Bartoli C, Rivera J, Tazi J, Guzmán G, Varela I, Depetris D, de Carlos F, Cobo J, Andrés V, Sandre-Giovannoli AD, Freije JM, Lévy N, López-Otín C. Splicing-Directed Therapy in a New Mouse Model of Human Accelerated Aging. Sci. Transl. Med. 2011;3:106ra107 doi:10.1126/scitranslmed.3002847
6 mg/Kg each of 2 sequences (or 12 mg/Kg control sequence) were administered to progeria model mice by tail vein injection twice per week for 12 weeks, starting at six weeks of age.
Reissner KJ, Sartor GC, Vazey EM, Dunn TE, Aston-Jones G, Kalivas PW. Use of vivo-morpholinos for control of protein expression in the adult rat brain. J Neurosci Meth. 2011;[Epub ahead of print] doi:10.1016/j.jneumeth.2011.10.009
Intracranial injections in rat brains of oligos targeting three different RNA targets, some though cannula and some by direct injection. Alternate URL here.
NOTE: Kate Reissner informs us that the doses reported in this paper in nmol should have been reported in pmol; that is, the reported doses are 1000x too high.
Shi YB, Hasebe T, Fu L, Fujimoto K, Ishizuya-Oka A. The development of the adult intestinal stem cells: Insights from studies on thyroid hormone-dependent amphibian metamorphosis. Cell Biosci. 2011 Sep 6;1(1):30.
Injection into stage 53/54 Xenopus tadpoles, knockdown in gut.
Zammarchi F, de Stanchina E, Bournazou E, Supakorndej T, Martires K, Riedel E, Corben AD, Bromberg JF, Cartegni L. Antitumorigenic potential of STAT3 alternative splicing modulation. Proc Natl Acad Sci U S A. 2011 Oct 17. [Epub ahead of print]
Injections of Vivo-Morpholinos in a mouse tumor model either intratumorally at a final concentration of 0.12 mg/30 μl or intravenously at 15 mg/kg. Oligos were administered 2x week for either 1 or 3 weeks.
Nakamura K, Du L, Tunuguntla R, Fike F, Cavalieri S, Morio T, Mizutani S, Brusco A, Gatti RA. Functional characterization and targeted correction of ATM mutations identified in Japanese patients with ataxia-telangiectasia. Hum. Mutat. 2011;[Epub ahead of print] doi:10.1002/humu.21632
Splice correction in lymphoblastoid cell line from patient.
Quinn M, Ueno Y, Pae HY, Huang L, Frampton G, Galindo C, Francis H, Horvat D, McMillin M, Demorrow S. Suppression of the HPA axis during extrahepatic biliary obstruction induces cholangiocyte proliferation in the rat. Am J Physiol Gastrointest Liver Physiol. 2012 Jan 1;302(1):G182-93. doi: 10.1152/ajpgi.00205.2011. Epub 2011 Oct 6.
Intracerebroventricular injection of Vivo-Morpholinos in rat, daily 4ug/rat/day for 7 days.
Taniguchi-Ikeda M, Kobayashi K, Kanagawa M, Yu CC, Mori K, Oda T, Kuga A, Kurahashi H, Akman HO, DiMauro S, Kaji R, Yokota T, Takeda S, Toda T. Pathogenic exon-trapping by SVA retrotransposon and rescue in Fukuyama muscular dystrophy. Nature. 2011;478:127-31 doi:10.1038/nature10456
FCMD model mice, human FCMD lymphoblasts and myotubes.
Vorlová S, Rocco G, LeFave CV, Jodelka FM, Hess K, Hastings ML, Henke H, Cartegni L. Induction of Antagonistic Soluble Decoy Receptor Tyrosine Kinases by Intronic PolyA Activation. Mol Cell. 2011;(43)6:927-39.
Cultured cells: HeLa, MDA-231 & HUVEC.
Nazmi A, Dutta K, Basu A. RIG-I Mediates Innate Immune Response in Mouse Neurons Following Japanese Encephalitis Virus Infection. PLoS ONE. 2011;6(6): e21761. doi:10.1371/journal.pone.0021761
Wu B, Benrashid E, Lu P, Cloer C, Zillmer A, Shaban M, Lu QL. Targeted Skipping of Human Dystrophin Exons in Transgenic Mouse Model Systemically for Antisense Drug Development. PLoS ONE. 2011;6(5):e19906. doi:10.1371/journal.pone.0019906
Kowalik L, Hudspeth AJ. A search for factors specifying tonotopy implicates DNER in hair-cell development in the chick's cochlea. Dev Biol. 2011 Jun 15;354(2):221-31. Epub 2011 Apr 8.
Vivo-Morpholinos locally injected in embryonic chick ears at E6.
Tsonis PA, Haynes T, Maki N, Nakamura K, Casco-Robles MM, Yamada S, Miura T, Chiba C, Del Rio-Tsonis K. Controlling gene loss of function in newts with emphasis on lens regeneration. Nature Protocols. 2011;6:593–599 doi:10.1038/nprot.2011.341 pdf
Widrick JJ, Jiang S, Choi SJ, Knuth ST, Morcos PA. An octaguanidine-morpholino oligo conjugate improves muscle function of mdx mice. Muscle & Nerve 2011 [Epub ahead of print]
Parra MK, Gee S, Mohandas N, Conboy JG. Efficient in vivo manipulation of alternative pre-mRNA splicing events using antisense morpholinos in mice. J Biol Chem. 2011 Feb 25;286(8):6033-9. Epub 2010 Dec 14.
Vivo-Morpholinos evaluated in mice at Lawrence Berkeley National Laboratory.
Nazmi A, Dutta K, Basu A. Antiviral and Neuroprotective Role of Octaguanidinium Dendrimer-Conjugated Morpholino Oligomers in Japanese Encephalitis. PLoS Negl Trop Dis 2010;4(11): e892. doi:10.1371/journal.pntd.0000892
Viral knockdowns in mice.
Carrillo M, Kim S, Rajpurohit SK, Kulkarni V, Jagadeeswaran P. Zebrafish von Willebrand factor. Blood Cells Mol Dis. 2010 Oct 28. [Epub ahead of print]
Vivo-Morpholinos in adult zebrafish.
Guo Y, Ma L, Cristofanilli M, Hart RP, Hao A, Schachner M. Transcription factor Sox11b is involved in spinal cord regeneration in adult zebrafish. Neuroscience. 2010 Oct 14. [Epub ahead of print]
Vivo-Morpholino on Gelfoam applied to zebrafish spinal cord transection at time of injury.
Kang JK, Malerba A, Popplewell L, Foster K, Dickson G. Antisense-induced Myostatin Exon Skipping Leads to Muscle Hypertrophy in Mice Following Octa guanidine Morpholino Oligomer Treatment. Mol Ther. 2010 Oct 5. [Epub ahead of print]
Matsuda H, Shi YB. An Essential and Evolutionarily Conserved Role of Protein Arginine Methyltransferase 1 for Adult Intestinal Stem Cells During Postembryonic Development. Stem Cells. 2010 Sep 16. [Epub ahead of print]
Intraperitoneal injection in Xenopus laevis.
Azoitei N, Pusapati GV, Kleger A, Möller P, Küfer R, Genze F, Wagner M, van Lint J, Carmeliet P, Adler G, Seufferlein T. Protein kinase D2 is a crucial regulator of tumour cell-endothelial cell communication in gastrointestinal tumours. Gut. 2010 Aug 23. [Epub ahead of print]
Vivo-Morpholinos in mice.
Vera T, Stec DE. Moderate Hyperbilirubinemia Improves Renal Hemodynamics in Angiotensin II Dependent Hypertension. Am J Physiol Regul Integr Comp Physiol. 2010 Oct;299(4):R1044-9. Epub 2010 Jul 28.
Vivo-Morpholinos in mice.
Kreidberg JA. WT1 and kidney progenitor cells. Organogenesis. 2010 Apr;6(2):61-70. Knockdown of WT1 in murine kidney explants; review offering perspective on experiments in Hartwig et al., Development 2010 (below).
Lecce L, Day M, Murphy CR. Abstract 15. ICAM-1 Is Involved in Uterine Receptivity in Rats. The Society for the Study of Reproduction 2010 Annual Meeting.
Maki N, Suetsugu-Maki R, Sano S, Nakamura K, Nishimura O, Tarui H, Del Rio-Tsonis K, Ohsumi K, Agata K, Tsonis PA. Oocyte-type linker histone B4 is required for transdifferentiation of somatic cells in vivo. FASEB J. 2010 Sep;24(9):3462-7. Epub 2010 May 11.
This describes Vivo-Morpholino use in a newt.
Presentations using Vivo-Morpholinos from 2010 American Society of Gene & Cell Therapy meeting:
See abstract 14 on this linked page.
See abstract 16 on this linked page.
Hartwig S, Ho J, Pandey P, Macisaac K, Taglienti M, Xiang M, Alterovitz G, Ramoni M, Fraenkel E, Kreidberg JA. Genomic characterization of Wilms' tumor suppressor 1 targets in nephron progenitor cells during kidney development. Development. 2010 Apr;137(7):1189-203.
Knockdown of WT1 in murine kidney explants.
Chablais F, Jazwinska A. IGF signaling between blastema and wound epidermis is required for fin regeneration. Development. 2010 Mar;137(6):871-9.
Injection of 1.5 mM Vivo-Morpholino into regenerating zebrafish fins.
Kim S, Radhakrishnan UP, Rajpurohit SK, Kulkarni V, Jagadeeswaran P. Vivo-Morpholino knockdown of alphaIIb: A novel approach to inhibit thrombocyte function in adult zebrafish. Blood Cells Mol Dis. 2010 Mar 15;44(3):169-74. Epub 2010 Jan 4.
Knockdown of alphaIIb in thrombocytes by i.v. injection of Vivo-Morpholinos into adult zebrafish.
Pérez B, Rincón A, Jorge-Finnigan A, Richard E, Merinero B, Ugarte M, Desviat LR. Pseudoexon exclusion by antisense therapy in methylmalonic aciduria (MMAuria). Hum Mutat. 2009 Sep 2. [Epub ahead of print]
Report of Vivo-Morpholinos compared with unmodified Morpholinos and Endo-Porter when tested in cultured human fibroblasts.
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.