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Knock down gadd45g


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#1 michalcn

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Posted 17 February 2011 - 08:32 AM

does anybody know how to knock down gadd45g ?
I know many of the downstream targets but in order to finish the prove that its gadd45g mediated I need to knock it down
the only sequence that I found working good based on the publication was mouse and I need human
I bought already 3 different si and sh with no successes
any suggestions?
I'm totally stuck now
thanks
Michal

#2 pcrman

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Posted 17 February 2011 - 06:43 PM

Do you have some siRNA against other genes and known to work? You can use them as positive control to give you an idea your system is OK such as transfection efficiency of your cell lines. Are your siRNAs still good without degradation?--run a simple agarose gel to verify that. If every thing is ok, you have to redesign new targets, although it is weird the all 3 siRNAs did not work.

#3 Jon Moulton

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Posted 18 February 2011 - 07:42 AM

Candal E, Thermes V, Joly JS, Bourrat F. Medaka as a model system for the characterisation of cell cycle regulators: a functional analysis of Ol-Gadd45gamma during early embryogenesis. Mech Dev. 2004 Jul;121(7-8):945-58.

It's been done in medaka fish using Morpholinos. They also work in cell culture when used with a delivery strategy (electroporation, Endo-Porter, etc.).

Open-access: .pdf article
Jon D. Moulton
Gene Tools, LLC
www.gene-tools.com

#4 michalcn

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Posted 23 February 2011 - 01:24 PM

thank you for the paper
but I'm looking at human samples so do you think that based on the homologies between the species I can use the same sequence to knock down human gadd45g?
Michal

#5 michalcn

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Posted 07 March 2011 - 07:26 AM

I previously asked how to knock down gadd45g I'm using shRNA without success and I still can't
you quote a paper using knock down in morpholino
but I'm looking at human samples so do you think that based on the homologies between the species I can use the same sequence to knock down human gadd45g?
do you know any other way of knocking down this gene maybe a drug?

sorry, but i don't know how to use the web site as continue the same question

Michal

#6 Jon Moulton

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Posted 07 March 2011 - 05:06 PM

Hi Michal,

You can continue a conversation in the same topic by adding a reply instead of opening a new topic (here I replied to your post).

The sequence needed is likely different, there has been plenty of sequence divergence.

If you contact us at Gene Tools, we'd be happy to discuss your target and suggest an oligo design based on your target gene (there is no charge for that). Feel free to email custsupport@gene-tools.com if you would like to explore the possibility of using Morpholinos. Dr. Alexandra Vincent is taking care of that email box; she can answer your questions about the mode of action of Morpholinos, design sequences and suggest strategies for your particular experimental problem. If you like you are also welcome to contact me directly at jmoulton@gene-tools.com

Kind regards,

- Jon
Jon D. Moulton
Gene Tools, LLC
www.gene-tools.com




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