Improved translation efficiency of injected mRNA during early embryonic development.
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.
Fink
Maria
M
Institute of Animal Breeding and Genetics, University of Veterinary Medicine, Vienna, Austria.
Flekna
Gabriele
G
Ludwig
Alfred
A
Heimbucher
Thomas
T
Czerny
Thomas
T
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D002678:Chimera / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005914:Globins / Q000235:genetics; D066298:In Vitro Techniques; D049409:Luciferases, Firefly / Q000235:genetics; D008845:Microinjections; D009990:Oryzias / Q000196:embryology* / Q000235:genetics* / Q000378:metabolism; D051762:PAX2 Transcription Factor / Q000235:genetics; D011061:Poly A / Q000235:genetics; D014176:Protein Biosynthesis; D020871:RNA Stability; D012333:RNA, Messenger / Q000008:administration & dosage / Q000235:genetics* / Q000378:metabolism; D020506:Untranslated Regions
D017931:DNA Primers; D051762:PAX2 Transcription Factor; D012333:RNA, Messenger; D020506:Untranslated Regions; D011061:Poly A; D005914:Globins; D049409:Luciferases, Firefly