2 replies to this topic

[altobarn]

Hi all,

I would like to perform down-regulation experiment in vitro by transfecting antagomir to block the production of endogenous Wnt1 protein. That means I would like to have a cell line which produce endogenous Wnt1 protein. Anyone out there has any clue if such cell lines exist?

Any ideas are welcome! Thanks.

-Altobarn-

[gfischer]

altobarn, on Aug 26 2009, 03:13 AM, said:

Hi all,

I would like to perform down-regulation experiment in vitro by transfecting antagomir to block the production of endogenous Wnt1 protein. That means I would like to have a cell line which produce endogenous Wnt1 protein. Anyone out there has any clue if such cell lines exist?

Any ideas are welcome! Thanks.

-Altobarn-

The simplest thing to do would be to look at what tissues Wnt1 is expressed in and get a cell line derived from that tissue type.  You could also transfect a cell line like HEK293 to express it, though this is a little more involved.

[altobarn]

Thanks for the suggestion. As I don't have any Wnt1 expressing plasmid, thus I would need extra time to do the cloning work and for this reason, I would prefer to skip this labourous task. One cell line came to my mind is SH-SY5Y. This neuroblastoma cell line are dopamine beta hydroxylase active. Given the fact that Wnt1 is required for dopaminergic neurons development, I speculate that SH-SY5Y might be having certain level of endogenous Wnt1. However I am hoping there is a better cell line by which the Wnt1 protein had been shown to be expressed  endogenously.

Any idea peps?