Synthesis of Functional mRNA in Mammalian Cells by Bacteriophage T3 RNA Polymerase
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Key result
The 5' nontranslated leader sequence from encephalomyocarditis virus allowed transcripts synthesized by T3 RNA polymerase in mammalian cells to be translated in a cap-independent fashion, yielding at least 10-fold higher expression than conventional vectors.
Population
Cultured mammalian cell lines (HeLa, BSC40, Ltk-, NIH 3T3, and C127)
Comparison
T3 RNA polymerase gene and EMCV-CAT fusion gene… vs Conventional transient expression vector…
Design
Preclinical
Authors
Discussion
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Implication
May aid mammalian gene expression studies; leaves open clinical or in vivo translation of cap-independent vectors.
Structured PICO
P
Population
Cultured mammalian cell lines (HeLa, BSC40, Ltk-, NIH 3T3, and C127)
I
Intervention
T3 RNA polymerase gene and EMCV-CAT fusion gene expression system
C
Comparator
Conventional transient expression vector dependent on host RNA polymerase II (pRSV-CAT)
O
Outcome
Level of gene expression (chloramphenicol acetyltransferase [CAT] activity)surrogate
The T3-EMCV expression system allows for high-level, cap-independent transient gene expression in a broad range of mammalian cell types.
Cite This Study
Zhou et al. (1990) studied this question. T3 RNA polymerase with EMCV leader sequence vs. Conventional transient expression vector (RNA polymerase II) was evaluated on Gene expression levels (CAT activity). The 5' nontranslated leader sequence from encephalomyocarditis virus allowed transcripts synthesized by T3 RNA polymerase in mammalian cells to be translated in a cap-independent fashion, yielding at least 10-fold higher expression than conventional vectors.
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