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Jon's Blog
| Post date | ||
|---|---|---|
| Morpholinos to validate target specificity of antibodies |
Here's an example of using Morpholinos to validate target specificity of antibodies. Belgacem YH, Borodinsky LN. Inversion of Sonic hedgehog action on its canonical pathway by electrical activity. Proc Natl Acad Sci U S A. 2015 Mar 31;112(13):4140-5. doi: 10.1073/pnas.1419690112. Epub 2015 Mar 17. |
Friday, June 12, 2015 - 10:54 |
| Websites (and some papers) about morphant and mutant comparisons |
I've been watching several websites with discussions of morphant and mutant comparisons. Here are links to the sites. The comments sections have been very supportive of Morpholino technology. I'll update this page as I find more. |
Thursday, March 5, 2015 - 13:23 |
| Morpholinos, mutants, and phenocopying |
I was concerned when Stefan Schulte-Merker and Didier Stainier' paper appeared in mid-2014, first in Development and then on The Node. Later in 2014 Nathan Lawson's paper in Developmental Cell was published. Prior to these papers I was intrigued and excited by CRISPR technology but felt that, while it might temporarily displace some use of Morpholinos, in the long run it would benefit Morpholino use by making the zebrafish model stronger. |
Wednesday, February 11, 2015 - 15:18 |
| Change in Morpholino activity over time |
Here are some notes discussing how Morpholino oligos can change activity over time, along with strategies for recovering and maintaining activity. Change over time in Morpholino activity |
Monday, January 5, 2015 - 10:59 |
| Xenopus laevis: brain ventricle injection then electroporation |
Faulkner RL, Wishard TJ, Thompson CK, Liu H-H, Cline HT. FMRP regulates neurogenesis in vivo in Xenopus laevis tadpoles. eNeuro. 2014;[Epub ahead of print] doi:10.1523/ENEURO.0055-14.2014 http://eneuro.org/content/eneuro/early/2014/12/31/ENEURO.0055-14.2014.fu... |
Friday, January 2, 2015 - 12:15 |
| How much Morpholino to inject into a zebrafish egg or early zygote? |
An injection volume of 500 pL is reasonable (Rosen et al. 2009, http://www.jove.com/video/1115/microinjection-of-zebrafish-embryos-to-an...). The basic rule for dosing of Morpholino injections is to use the least dose that gives you the phenotype of interest. As the dose increases, so does the potential for off-target RNA interactions and, with that, the potential of seeing a phenotype that has nothing to do with your target RNA. |
Wednesday, December 31, 2014 - 08:21 |
| Intestinal delivery to zebrafish from fish water |
Progatzky F, Sangha NJ, Yoshida N, McBrien M, Cheung J, Shia A, Scott J, Marchesi JR, Lamb JR, Bugeon L, Dallman MJ. Dietary cholesterol directly induces acute inflammasome-dependent intestinal inflammation. Nat Commun. 2014 Dec 23;5:5864. doi: 10.1038/ncomms6864. http://www.nature.com/ncomms/2014/141223/ncomms6864/full/ncomms6864.html This paper claims delivery of Morpholino to the intestinal epithelium of zebrafish by putting Morpholinos in the fish water around feeding time. |
Friday, December 26, 2014 - 13:17 |
| Rescues with wild-type or mutant mRNA |
Here is an interesting strategy for nailing down the relationship between a phenotype and a particular mutation. Using a zebrafish pnpla6 morphant, co-injection of wild-type human pnpla6 mRNA rescued the morphant phenotype while mutant human pnpla6 mRNA failed to rescue. |
Wednesday, December 10, 2014 - 13:54 |
| Of Mice and Morpholinos, 2014 |
These are some papers using Morpholinos in mouse, published in 2014. Vivo-Morpholino WT1 controls antagonistic FGF and BMP-pSMAD pathways in early renal progenitors . Motamedi FJ, Badro DA, Clarkson M, Rita Lecca M, Bradford ST, Buske FA, Saar K, Hübner N, Brändli AW, Schedl A. Nat Commun. 2014 Jul 17;5:4444. doi: 10.1038/ncomms5444. |
Tuesday, October 28, 2014 - 11:12 |
| Some Morpholino Citations for Hematology: 2014 |
Representative Recent Morpholino Citations for Hematology: 2014 Characterization of an apparently synonymous F5 mutation causing aberrant splicing and factor V deficiency. |
Tuesday, October 21, 2014 - 13:34 |
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