Factors limiting autogene-based cytoplasmic expression systems Jonathan Finn * ,1 , Ian MacLachlan † and Pieter Cullis * ,‡ * Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada; † Protiva Biotherapeutics Inc., Burnaby, British Columbia, Canada; and ‡ Inex Pharmaceuticals Corp., Burnaby, British Columbia, Canada 1 Correspondence: University of British Columbia, Dept. of Biochemistry and Molecular Biology, 2146 Health Sciences Mall, Vancouver, BC V6T 1Z3, Canada. E-mail: jdfinn@interchange.ubc.ca <h3>SPECIFIC AIMS</h3> The purpose of this study was to determine the factors limiting the autogene-based cytoplasmic expression system and determine ways in which levels of gene expression can be increased. 1) The CMV promoter was replaced with the RSV or SV40 promoter and its effect on nuclear and cytoplasmic expression was examined; 2) The T7 phage RNA polymerase (RNAP) gene was replaced and compared with SP6 or T3 RNAP genes; 3) The EMCV internal ribosome entry site (IRES) currently used to allow for the translation of uncapped cytoplasmic transcripts was replaced and compared with eIF4G or Gtx IRES elements; 4) Translation efficiency of capped, uncapped, or EMCV IRES containing mRNA transcripts was compared to determine the efficiency of the EMCV IRES at driving translation;
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Factors limiting autogene-based cytoplasmic expression systems
Finn, Jonathan; MacLachlan, Ian; Cullis, Pieter
Follow The FASEB Journal
, Volume 19 (6): 608
Fed of American Socs for Experimental Biology – Apr 1, 2005
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