The study demonstrates that spurious splicing of the XIAP 5' UTR is an artifact specific to the Rluc/Fluc reporter system, which is important for accurately studying its internal ribosome entry site (IRES) activity.
Urges caution interpreting XIAP IRES data from Rluc/Fluc reporters; leaves open optimal bicistronic validation methods.
X-chromosome-linked inhibitor of apoptosis, XIAP, has been shown to contain a strong internal ribosome entry site (IRES) within its 5' untranslated region (UTR) that promotes translation of XIAP mRNA under conditions of cellular stress. This claim came under scrutiny in a recent report demonstrating that the XIAP 5' UTR undergoes splicing when inserted between the two reporter cistrons of the dual luciferase plasmid Rluc/Fluc. In this paper, we demonstrate that the splicing within the XIAP 5' UTR specifically occurs only in the context of mRNA produced from the Rluc/Fluc but not the pbetagal/CAT bicistronic reporter plasmid.
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Holčı́k et al. (2005) studied this question.
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