RNA Interference in Pigs: Comparison of RNAi Test Systems and Expression Vectors |
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Authors: | Claudia Merkl Simon Leuchs Anja Saalfrank Alexander Kind Angelika Schnieke |
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Institution: | (1) Chair of Livestock Biotechnology, TU Muenchen, Liesel-Beckmann-Str. 1, 85354 Freising, Germany; |
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Abstract: | We have examined the use of RNA interference as a means of downregulating gene expression and provide the first comparison
of shRNA and artificial miRNA constructs for transgenic livestock. Several in vitro assays were performed to identify the
most effective RNAi constructs. shRNA and miRNA constructs achieved significant downregulation of two porcine target genes:
the milk whey protein beta-lactoglobulin and the tumour suppressor p53. Results of different assays were, however, sometimes
at variance, indicating that no one assay can be relied upon to predict the effectiveness of an RNAi construct. Our findings
are that screening of RNAi constructs is most informative if carried out in primary cells that express the target gene and
are competent for somatic cell nuclear transfer. Importantly, the use of miRNA constructs makes tissue specific gene knockdown
in large animals a realistic possibility. |
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Keywords: | |
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