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Morpholino Oligos:
   Making Sense of Antisense?
                   Based on the review of Janet Heasman
             Divi...
Overview
• Downregulating gene expression helps
  understanding their functions
• Most of the experiments are done to unde...
Structures




             Genie Genetique – Sebastian König and
22.05.2009
                       Sebastian Olényi
Principle




                               “DGI reliably
                               achieves >90%
                  ...
Advantages
• Equal or higher efficacy and specificity
  greatly superior to any other antisense
  structural type
• Morpho...
Disadvantages
• Completely artificial molecule, so no
  vectors possible
• Delivery usually only via microinjection or
  e...
Application
• 25mers, Little self-complementarity (less than 4
  contiguous intrastrand base pairs)
• Less than 36% guanin...
Genie Genetique – Sebastian König and
22.05.2009
                       Sebastian Olényi
Thank you!

                                                      References
http://www.sanger.ac.uk/Info/Press/gfx/zebraf...
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Morpholino Oligos

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Morpholino Oligos are an interesting technique to silence Genes. Learn more about their usage and potential.

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Morpholino Oligos

  1. 1. Morpholino Oligos: Making Sense of Antisense? Based on the review of Janet Heasman Division of Developmental Biology, Children’s Hospital Medical Center, Cincinnati, Ohio Developmental Biology 243, 209–214 (2002) 22.05.2009 Genie Genetique – Sebastian König and Sebastian Olényi
  2. 2. Overview • Downregulating gene expression helps understanding their functions • Most of the experiments are done to understand embryonic development • Morpholinos are synthetic, usually 25 bases long, with a different sugar-structure • Typical model organisms: zebrafish, African clawed frog, sea urchin, chick and mice. Genie Genetique – Sebastian König and 22.05.2009 Sebastian Olényi
  3. 3. Structures Genie Genetique – Sebastian König and 22.05.2009 Sebastian Olényi
  4. 4. Principle “DGI reliably achieves >90% knockdown of gene expression” Genie Genetique – Sebastian König and 22.05.2009 Sebastian Olényi
  5. 5. Advantages • Equal or higher efficacy and specificity greatly superior to any other antisense structural type • Morpholinos do not activate the complement cascade or the interferon system and are not degraded • Excellent solubility and stability • Working at high concentrations is possible • No need for RNase H to ensure their activity Genie Genetique – Sebastian König and 22.05.2009 Sebastian Olényi
  6. 6. Disadvantages • Completely artificial molecule, so no vectors possible • Delivery usually only via microinjection or electroporation • Not always high effects, uncertain amount of downregulation • Side-effects for high dosage, oligo- dependent Genie Genetique – Sebastian König and 22.05.2009 Sebastian Olényi
  7. 7. Application • 25mers, Little self-complementarity (less than 4 contiguous intrastrand base pairs) • Less than 36% guanine content • No runs of more than triplets of GGG • Concentrations between 1 and 12μM • 2-4 days effectiveness • Controls with – mRNA rescue experiments – Two different morpholinos directed to different gene parts with same effect – Other over- and/or underexpression studies Genie Genetique – Sebastian König and 22.05.2009 Sebastian Olényi
  8. 8. Genie Genetique – Sebastian König and 22.05.2009 Sebastian Olényi
  9. 9. Thank you! References http://www.sanger.ac.uk/Info/Press/gfx/zebrafish.jpg http://www.columbia.edu/itc/cerc/danoff-burg/invasion_bio/inv_spp_summ/xenopuslaevis.jpg http://www.alaskaunderseatours.com/photos/Red-Sea-Urchin.jpg http://www.gene-tools.com/products_and_applications http://www.bentham.org/ctmc/openaccessarticles/ctmc7-7/0003R.pdf Genie Genetique – Sebastian König and 22.05.2009 Sebastian Olényi

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