mRNA with a <20-nt poly(A) tail imparted by the poly(A)-limiting element is translated as efficiently in vivo as long poly(A) mRNA

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FIGURE 1.
FIGURE 1.

The PLE imparts a short poly(A) tail on luciferase mRNA. The sequence corresponding to PLE B (shown above the figure) was inserted downstream of the firefly luciferase gene, 12 bp upstream of a minimal synthetic polyadenylation element (SPA; Das Gupta et al. 1998) in a plasmid with transcription controlled by the CMV promoter. Poly(A) tail length of mRNA expressed in transiently transfected cells was determined by PAT assay (Salles et al. 1999) using an oligo(dT) primer-adapter to prime reverse transcription and a 5′ [32P]-labeled upstream primer for luciferase. The amplified products were separated on a denaturing 6% polyacrylamide/urea gel and visualized by PhosphorImager analysis. The lane marked M contains size markers of [32P]-labeled HinfI digested [phis]X174 DNA.

This Article

  1. Published in Advance May 31, 2005, doi: 10.1261/rna.2470905 RNA 11: 1131-1140

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