I have a phenotype, but northern says gene is not knocked down - rnai knockdown (Apr/03/2005 )
I have recevied a line that is an rnai knockdown (supposedly) of a histone gene. I am getting a phenotype from this line that is segregating with the construct, however my northern analysis tells me that the endogenous gene is expressing rna just fine and is therefore not knocked down. I do not think the generic construct is causing the phenotype, since construct controls without the rnai sequence do not exhibit the phenotype.
How can I get rna expression but still have a phenotype? Should I do a western?
Thanks.
-transposer-
QUOTE (transposer @ Apr 3 2005, 12:59 PM)
I have recevied a line that is an rnai knockdown (supposedly) of a histone gene. I am getting a phenotype from this line that is segregating with the construct, however my northern analysis tells me that the endogenous gene is expressing rna just fine and is therefore not knocked down. I do not think the generic construct is causing the phenotype, since construct controls without the rnai sequence do not exhibit the phenotype.
How can I get rna expression but still have a phenotype? Should I do a western?
Thanks.
How can I get rna expression but still have a phenotype? Should I do a western?
Thanks.
Hi!
It is indeed quite common to see antisense or siRNA effects only on translational levels and not on the RNA levels.
You should do a western if possible to confirms this.
regards, Søren
Søren M. Echwald, MSc., Ph.D.
-----------------------------
Exiqon A/S
Bygstubben 9
DK-2950 Vedbaek
Denmark
www.Exiqon.com
-smesme-
hi
definitely do a western blot.
but you can also do in a parallel experiment a northern to see the presence of your siRNA.
-fred_33-