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)

05/22/09 Genie Genetique – Sebastian König

and Sebastian Olényi
 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.

05/22/09 Genie Genetique – Sebastian König

and Sebastian Olényi
 Structures

05/22/09 Genie Genetique – Sebastian König

and Sebastian Olényi
 Principle

“DGI reliably
achieves
>90%
knockdown of
gene
Genie Genetique – Sebastian König
05/22/09
and Sebastian Olényi
 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

05/22/09 Genie Genetique – Sebastian König

and Sebastian Olényi
 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

05/22/09 Genie Genetique – Sebastian König

and Sebastian Olényi
 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

05/22/09 Genie Genetique – Sebastian König

and Sebastian Olényi
05/22/09 Genie Genetique – Sebastian König
and Sebastian Olényi
 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

05/22/09 Genie Genetique – Sebastian König

and Sebastian Olényi