Improved translation efficiency of injected mRNA during early embryonic development

被引：26

作者：

Fink, Maria ^{[1
]}

Flekna, Gabriele ^{[1
]}

Ludwig, Alfred ^{[1
]}

Heimbucher, Thomas ^{[1
]}

Czerny, Thomas ^{[1
]}

机构：

[1] Vet Univ Vienna, Inst Anim Breeding & Genet, A-1210 Vienna, Austria

来源：

DEVELOPMENTAL DYNAMICS | 2006年 / 235卷 / 12期

关键词：

mRNA injection; UTR; beta-globin; SV40 late poly(A); T7TS; pCS2; pMC;

D O I：

10.1002/dvdy.20995

中图分类号：

R602 [外科病理学、解剖学]; R32 [人体形态学];

学科分类号：

100101 ;

摘要：

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.

引用

页码：3370 / 3378

页数：9

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