Beta-thalassemia treated in mice with antisense - Morpholino oligos block mutant splice site, restore wild-type splicing (Feb/06/2009 )

Morpholinos conjugated with cell-penetrating peptides were injected into mice carrying a splice-site mutation in beta-globin. The Morpholinos blocked the mutant splice sites and restored normal splicing so that the mice produced fully-functional hemoglobin.

Svasti S, Suwanmanee T, Fucharoen S, Moulton HM, Nelson MH, Maeda N, Smithies O, Kole R. RNA repair restores hemoglobin expression in IVS2-654 thalassemic mice. Proc Natl Acad Sci U S A. 2009 Jan 27;106(4):1205-10. Epub 2009 Jan 21.

http://www.ncbi.nlm.nih.gov/pubmed/19164558

Jon Moulton on Feb 6 2009, 09:34 AM said:

That surely sounds novel.

I was not aware of anybody having successfully used morpholino in mice, let also therapeutically. But I am not much into this field.

i just want to ask because i'm so curios about it how genes affect in humans life???Hope you will answer all my question......

cellcounter on Feb 6 2009, 07:50 AM said:

They have been used a fair amount in mouse embryos by microinjecting oocytes. Their use in adult mice is more recent. The peptide-conjugated Morpholinos have been used in virology studies in adult mice and in development of a therapeutic for Duchenne muscular dystrophy (DMD). Vivo-Morpholinos, linked to an octaguanidinium dendrimer rather than a cell-penetrating peptide, are a more recent in vivo delivery strategy.

This citation list is not comprehensive. I've cited a few papers in each category.

In mouse embryos:

Coonrod SA, Bolling LC, Wright PW, Visconti PE, Herr JC. A morpholino phenocopy of the mouse MOS mutation. Genesis. 2001 Jul;30(3):198-200.

Mellitzer G, Hallonet M, Chen L, Ang S. Spatial and temporal ‘knock down’ of gene expression by electroporation of double-stranded RNA and morpholinos into early postimplantation mouse embryos. Mech Dev. 2002 Oct;118(1-2):57.

Siddall LS, Barcroft LC, Watson AJ. Targeting gene expression in the preimplantation mouse embryo using morpholino antisense oligonucleotides. Mol Reprod Dev. 2002 Dec;63(4):413-421.

Kanzler B, Haas-Assenbaum A, Haas I, Morawiec L, Huber E, Boehm T. Morpholino oligonucleotide-triggered knockdown reveals a role for maternal E-cadherin during early mouse development. Mech Dev. 2003 Dec;120(12):1423-1432.

Cheng TC, Huang CC, Chen CI, Liu CH, Hsieh YS, Huang CY, Lee MS, Liu JY. Leukemia Inhibitory Factor Antisense Oligonucleotide Inhibits the Development of Murine Embryos at Preimplantation Stages. Biol Reprod. 2004 May;70(5):1270-6. Epub 2003 Dec 26.

Homer HA, McDougall A, Levasseur M, Yallop K, Murdoch AP, Herbert M. Mad2 prevents aneuploidy and premature proteolysis of cyclin B and securin during meiosis I in mouse oocytes. Genes Dev. 2005 Jan 15;19(2):202-7.

Reis A, Chang HY, Levasseur M, Jones KT. APCcdh1 activity in mouse oocytes prevents entry into the first meiotic division. Nat Cell Biol. 2006 May;8(5):539-40.

Reis A, Madgwick S, Chang HY, Nabti I, Levasseur M, Jones KT. Prometaphase APCcdh1 activity prevents non-disjunction in mammalian oocytes. Nat Cell Biol. 2007 Oct;9(10):1192-8. Epub 2007 Sep 23.

Foygel K, Choi B, Jun S, Leong DE, Lee A, Wong CC, Zuo E, Eckart M, Reijo Pera RA, Wong WH, Yao MW. A novel and critical role for Oct4 as a regulator of the maternal-embryonic transition. PLoS ONE. 2008;3(12):e4109. Epub 2008 Dec 31.

In adult mice:

McCaffrey AP, Meuse L, Karimi M, Contag CH, Kay MA. A potent and specific morpholino antisense inhibitor of hepatitis C translation in mice. Hepatology. 2003 Aug;38(2):503-8.

Warfield KL, Swenson DL, Olinger GG, Nichols DK, Pratt WD, Blouch R, Stein DA, Aman MJ, Iversen PL, Bavari S. Gene-Specific Countermeasures against Ebola Virus Based on Antisense Phosphorodiamidate Morpholino Oligomers. PLoS Pathog. 2006 Jan;2(1):e1. Epub 2006 Jan 13.

Stein DA. Inhibition of RNA virus infections with peptide-conjugated morpholino oligomers. Curr Pharm Des. 2008;14(25):2619-34.

For DMD studies in mice:

Fall AM, Johnsen R, Honeyman K, Iversen P, Fletcher S, Wilton SD. Induction of revertant fibres in the mdx mouse using antisense oligonucleotides. Genet Vaccines Ther. 2006 May 24;4:3.

Fletcher S, Honeyman K, Fall AM, Harding PL, Johnsen RD, Steinhaus JP, Moulton HM, Iversen PL, Wilton SD. Morpholino Oligomer-Mediated Exon Skipping Averts the Onset of Dystrophic Pathology in the mdx Mouse. Mol Ther. 2007 Sep;15(9):1587-92. Epub 2007 Jun 19.

Doran P, Wilton SD, Fletcher S, Ohlendieck K. Proteomic profiling of antisense-induced exon skipping reveals reversal of pathobiochemical abnormalities in dystrophic mdx diaphragm. Proteomics. 2009 Jan 8.

For Vivo-Morpholinos:

Morcos PA, Li YF, Jiang S. 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.

-Jon Moulton-

miechel on Feb 9 2009, 01:02 AM said:

i just want to ask because i'm so curios about it how genes affect in humans life???Hope you will answer all my question......

Hi Miechel,

Genes provide instructions your cells use to make proteins. Without genes, you would have no proteins and you would not live. So, genes critically affect human life, in fact they make all life of the kind we know possible. Anything we recognize as "living" has genes -- bacteria, fungi, protists, plants and animals all rely on genes to make proteins. Even viruses have genes, though they might not carry them in DNA (genes coded in RNA is an option some kinds of viruses use).

-Jon Moulton-

Jon Moulton on Feb 6 2009, 11:34 PM said:

Morpholinos conjugated with cell-penetrating peptides were injected into mice carrying a splice-site mutation in beta-globin. The Morpholinos blocked the mutant splice sites and restored normal splicing so that the mice produced fully-functional hemoglobin.

Svasti S, Suwanmanee T, Fucharoen S, Moulton HM, Nelson MH, Maeda N, Smithies O, Kole R. RNA repair restores hemoglobin expression in IVS2-654 thalassemic mice. Proc Natl Acad Sci U S A. 2009 Jan 27;106(4):1205-10. Epub 2009 Jan 21.

http://www.ncbi.nlm.nih.gov/pubmed/19164558

Hi Jon,

Long time we have not contacted each other. Since this website was updated several months ago and Biospace can not be accessed now, I could not get your contact information.

Because of these problems, I do not have any clue when I have questions about Morpholinos.

According to the Morpholino, I think it could be better researched during the embryogenesis. But how could I analyze the next generation?

Regards
Jixiang Kong

-stone757-

Hi Jon,

Long time we have not contacted each other. Since this website was updated several months ago and Biospace can not be accessed now, I could not get your contact information.

Because of these problems, I do not have any clue when I have questions about Morpholinos.

According to the Morpholino, I think it could be better researched during the embryogenesis. But how could I analyze the next generation?

Regards
Jixiang Kong

Hi Jixiang,

We now have modified Morpholinos that can be used in adult animals, so you can do knockdowns in later stage organisms. However, we've not solved the problem of delivery into the cytosol of plant cells, so in most cases I don't think Morpholinos will be useful in your system. Possibly injection into plant embryos could work, but the method has not been worded out. The effects of Morpholinos last form days to months, depending on the system, but I doubt that doing a knockdown and seeing the knockdown in the next generation will be a practical experimental approach (except perhaps for simple and rapidly-dividing organisms).

Good to hear from you. You can contact me at Gene Tools at the email address jmoulton <at> gene-tools.com, replacing <at> with @.

Best regards,

- Jon

-Jon Moulton-

Jon Moulton on Mar 10 2009, 02:45 AM said:

Hi Jon,

Long time we have not contacted each other. Since this website was updated several months ago and Biospace can not be accessed now, I could not get your contact information.

Because of these problems, I do not have any clue when I have questions about Morpholinos.

According to the Morpholino, I think it could be better researched during the embryogenesis. But how could I analyze the next generation?

Regards
Jixiang Kong

Hi Jixiang,

We now have modified Morpholinos that can be used in adult animals, so you can do knockdowns in later stage organisms. However, we've not solved the problem of delivery into the cytosol of plant cells, so in most cases I don't think Morpholinos will be useful in your system. Possibly injection into plant embryos could work, but the method has not been worded out. The effects of Morpholinos last form days to months, depending on the system, but I doubt that doing a knockdown and seeing the knockdown in the next generation will be a practical experimental approach (except perhaps for simple and rapidly-dividing organisms).

Good to hear from you. You can contact me at Gene Tools at the email address jmoulton <at> gene-tools.com, replacing <at> with @.

Best regards,

- Jon

I doubt this if this is from a novel. How come when directing pass to the genes.

-ichatfilipina-

I doubt this if this is from a novel. How come when directing pass to the genes.

Hi ichatfilipina,

Take a look at this abstract, it will explain more.

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. advance online publication, March 10, 2009
http://www.nature.com/mt/journal/vaop/ncur.../mt200938a.html

-Jon Moulton-

Jon Moulton on Mar 10 2009, 02:45 AM said:

Hi Jon,

Long time we have not contacted each other. Since this website was updated several months ago and Biospace can not be accessed now, I could not get your contact information.

Because of these problems, I do not have any clue when I have questions about Morpholinos.

According to the Morpholino, I think it could be better researched during the embryogenesis. But how could I analyze the next generation?

Regards
Jixiang Kong

Hi Jixiang,

We now have modified Morpholinos that can be used in adult animals, so you can do knockdowns in later stage organisms. However, we've not solved the problem of delivery into the cytosol of plant cells, so in most cases I don't think Morpholinos will be useful in your system. Possibly injection into plant embryos could work, but the method has not been worded out. The effects of Morpholinos last form days to months, depending on the system, but I doubt that doing a knockdown and seeing the knockdown in the next generation will be a practical experimental approach (except perhaps for simple and rapidly-dividing organisms).

Good to hear from you. You can contact me at Gene Tools at the email address jmoulton <at> gene-tools.com, replacing <at> with @.

Best regards,

- Jon

Hi Jon,

It is great that Morpholinos can be various modified and improved in some characteristics. and I think it will be used in plant systems in the future.

Recently I am busy with my thesis and application for Ph.D study in Europe for the embryogenesis, and I hope I would do some research on the using of morpholinos within the plant embryos.

And Also thanks for your email address.

Kind Regards
Jixiang

-stone757-

Pages: 1 2 Next