[Zbrafish] Low efficacy of splice-blocking morpholino

[Burdine, Rebecca D]

There may be maternal RNA in your embryos which your splice-blocking
morpholinos may not affect.

You don't say if you get a phenotype or not?  If you do, perhaps you can
use both morpholinos together at lower doses to enhance the penetrance.

If you aren't getting a phenotype, you may not be able to induce one or
you may need to knockdown maternal message with an ATG or UTR
morpholino.

Becky

---------------------------------------------------
Rebecca D. Burdine, Ph.D.
Assistant Professor
Dept. of Molecular Biology
Princeton University
Washington Road Mof 433
Princeton, NJ 08544 
 
Phone: (609) 258-7515
Fax: (609) 258-1343
Email: rburdine from princeton.edu
Admin Assistant: Cathy Falk (609) 258-1604

-----Original Message-----
From: zbrafish-bounces from oat.bio.indiana.edu
[mailto:zbrafish-bounces from oat.bio.indiana.edu] On Behalf Of
weibinz from umich.edu
Sent: Saturday, March 10, 2007 5:39 PM
To: Zbrafish from magpie.bio.indiana.edu
Subject: [Zbrafish] Low efficacy of splice-blocking morpholino

Hi,
I am testing two splicing -blocking morpholino rencently. I found the 
efficacy of blocking splicing is not high enough. I always detected the 
normaly processed mRNA in the embryos injected with morpholino, in the 
presence of the abberently processed mRNA due to the morpholino. My 
injected amount of morpholino is estimated to be about 16ng per
zebrafish 
embryo, which I think has hit the upper limit and some embryos( about
10%) 
already showed massive necrosis. I used either Danieau's or water to
dilute 
the morpholino to 0.5mM concentration and inject 2-4 nl per embryo. I am

wondering whether anybody could share with me some tips to increase the 
efficacy.

Thanks a lot.

weibin

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