3 replies to this topic

[Jaya Rantham]

Hi,

Can any one help me in giving me the protocol and suggestions to do gene silencing using SiRNA in brain organotypic slices of 350um grown on inserts with 1.1ml of normal growth media.

Thanks
Jaya.

Edited by Jaya Rantham, 19 May 2009 - 02:19 PM.

[Jon Moulton]

Jaya Rantham, on May 19 2009, 03:16 PM, said:

Hi,

Can any one help me in giving me the protocol and suggestions to do gene silencing using SiRNA in brain organotypic slices of 350um grown on inserts with 1.1ml of normal growth media.

Thanks
Jaya.

If you insist on doing the knockdown with siRNA I can't help.  Here is a citation for a paper using knockdown by unmodified Morpholinos and delivery by Endo-Porter in kidney organotypic cultures.  The same system should work in brain slices.

Nikopoulos G, Adams T, Adams D, Oxburgh L, Prudovsky I, Verdi J.  The use of Endo-Porter to deliver morpholinos in kidney organ culture.  Biotechniques. 2008 Apr;44(4):547-9.

[Jaya Rantham]

Thanks for the information Jon Moulton.

Jon Moulton, on May 19 2009, 05:51 PM, said:

Jaya Rantham, on May 19 2009, 03:16 PM, said:

Hi,

Can any one help me in giving me the protocol and suggestions to do gene silencing using SiRNA in brain organotypic slices of 350um grown on inserts with 1.1ml of normal growth media.

Thanks
Jaya.

If you insist on doing the knockdown with siRNA I can't help.  Here is a citation for a paper using knockdown by unmodified Morpholinos and delivery by Endo-Porter in kidney organotypic cultures.  The same system should work in brain slices.

Nikopoulos G, Adams T, Adams D, Oxburgh L, Prudovsky I, Verdi J.  The use of Endo-Porter to deliver morpholinos in kidney organ culture.  Biotechniques. 2008 Apr;44(4):547-9.

[rantes]

Hey Jaya.

I'm new to the forum and interested too in siRNA treatment in organotypic hippocampal cultures.

Did you find any new information about it?

I have found one article that uses this method  (JOURNAL OF NEUROCHEMISTRY | 2008 | 106 | 978–988).

Please let me know if it is any help and if you have any other info.

Thanks

Ran