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Improved translation efficiency of injected mRNA during early embryonic development
Author(s) -
Fink Maria,
Flekna Gabriele,
Ludwig Alfred,
Heimbucher Thomas,
Czerny Thomas
Publication year - 2006
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.20995
Subject(s) - biology , polyadenylation , messenger rna , untranslated region , coding region , translational efficiency , translation (biology) , transcription (linguistics) , microbiology and biotechnology , gene , three prime untranslated region , genetics , linguistics , philosophy
Injection techniques are a powerful approach to study gene function in fish and frog model systems. In particular, in vitro transcribed mRNA is broadly used for such misexpression experiments. Sequence elements flanking the coding region, such as untranslated repeats and polyadenylation sequences, are known to affect the stability and the translation efficiency of mRNA. Here we show that in early embryos, poly(A) signals strongly contribute to the activity of the injected mRNA. Of interest, they only marginally affect mRNA stability, whereas the translation efficiency is dramatically enhanced. Combination of a poly(A) tail and an SV40 late poly(A) signal leads to highly synergistic effects of the two elements for injected mRNA. Compared with established vector systems, we detected a 20‐fold improvement for mRNA derived from the novel transcription vector pMC. Developmental Dynamics 235:3370–3378, 2006. © 2006 Wiley‐Liss, Inc.