Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
- - - - -

Silencing of the first cistron

  • Please log in to reply
1 reply to this topic

#1 PurpleKnight



  • Active Members
  • Pip
  • 11 posts

Posted 26 February 2010 - 05:05 PM

Hi there,

Our lab uses a bicistronic vector linking our gene of interest (cistron 1) to luciferase (cistron 2) with an ECMV-IRES. Expression is driven by CMV promoter.
We use luciferase expression through monitoring of bioluminescence both in vitro and in vivo as a surrogate for monitoring expression of cistron 1.
While our cells are still bioluminescent, it seems that cistron 1' expression has decreased dramatically.
My PI is now telling me that there can be silencing of the first cistron (at the transcriptional level) with the IRES still promoting luciferase transcription, hence the bioluminescence observed... I am afraid she may have misunderstood the person who told her that but I just wanted to double check in case I was missing something.
Any input or advices would be welcome!
Thanks a bunch!

#2 Rsm


    Post Dog

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 385 posts

Posted 26 February 2010 - 08:18 PM


IRES-dependent reporter gene expression is not stoichiometric to first gene expression (see for example PMID 10933956). It depends on both first and second gene themselves, I had higher expression of IRES-eGFP than of the first gene. If you want to quantify first gene expression to reporter gene expression, I'd recommend using a 2A system. Here's some reference: http://www.virologyj.../content/3/1/14 (PMID 16539700).



Edited by Minna, 26 February 2010 - 08:31 PM.

I got soul, but I'm not a soldier

Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.