Regulated expression

Jose de las Heras josenet at tiscali.co.uk
Tue Mar 7 17:27:39 EST 2006


Doh!
You're absolutely right, of course... (whops... what an oversight)

I guess one could try another inducible approach, to induce the gene of 
interest and at the same time the appropriate shRNA against the gene we want 
to turn off. The problem may be that complete repression may be difficult to 
achieve. A while ago I was looking into making an RNA pol inducible promoter 
for a very similar purpose... however it all remained in the drawing board. 
But I am sure there must be inducible shRNA promoters around by now.

Jose

"Cornelius Fritsch" <C.Fritsch at dkfz-heidelberg.de> wrote in message 
news:op.s5zm68kry4bjlh at anjacorni...
> Isn't the difference between Tet-On and Tet-Off in the regulator? Which 
> would mean that in the presence of both negative and positive regulator 
> you have expression from BOTH promoters in the presence OR absence of 
> Dox...
>
> On Sat, 04 Mar 2006 00:51:38 +0100, Jose de las Heras 
> <josenet at tiscali.co.uk> wrote:
>
>>
>> <v.gukassyan at gmail.com> wrote in message
>> news:1141261734.787077.150550 at i40g2000cwc.googlegroups.com...
>>> Dear Colleagues,
>>>
>>> I'm looking for a way to regulate expression of two proteins
>>> simultaneously. After transfection I need expression of only one of two
>>> proteins of interest. After a while expression of the another should be
>>> launched witth the first one blocked at the same time.
>>>
>>> Will there be any suggestions on this specific regulation mechanism
>>> development? There is no difference at which level to conduct the
>>> regulation - transcription/translation, etc...
>>>
>>> Thank you in advance,
>>> Vladimir
>>
>> How about using tet-ON/OFF or another similar system?
>>
>> if teh first gene is on a tet-OFF promoter and the second on a tet-ON, 
>> you
>> transfect without doxycyclin and only the first gene will be expressed. 
>> Then
>> add doxycyclin when you want to make the switch, and the first one will 
>> be
>> repressed, and teh second activated at the same time.
>> Providing there's not much leakage this could work?
>>
>> JOse
>>
>>
>
>
>
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