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An RNAi-based approach to down-regulate a gene family in vivo

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Item Type:Article
Title:An RNAi-based approach to down-regulate a gene family in vivo
Creators Name:Kim, J. and Badaloni, A. and Willert, T. and Zimber-Strobl, U. and Kühn, R. and Wurst, W. and Kieslinger, M.
Abstract:Genetic redundancy poses a major problem to the analysis of gene function. RNA interference allows the down-regulation of several genes simultaneously, offering the possibility to overcome genetic redundancy, something not easily achieved with traditional genetic approaches. Previously we have used a polycistronic miR155-based framework to knockdown expression of three genes of the early B cell factor family in cultured cells. Here we develop the system further by generating transgenic mice expressing the RNAi construct in vivo in an inducible manner. Expression of the transgene from the strong CAG promoter is compatible with a normal function of the basal miRNA/RNAi machinery, and the miR155 framework readily allows inducible expression from the Rosa26 locus as shown by Gfp. However, expression of the transgene in hematopoietic cells does not lead to changes in B cell development and neuronal expression does not affect cerebellar architecture as predicted from genetic deletion studies. Protein as well as mRNA levels generated from Ebf genes in hetero- and homozygous animals are comparable to wild-type levels. A likely explanation for the discrepancy in the effectiveness of the RNAi construct between cultured cells and transgenic animals lies in the efficiency of the sequences used, possibly together with the complexity of the transgene. Since new approaches allow to overcome efficiency problems of RNAi sequences, the data lay the foundation for future work on the simultaneous knockdown of several genes in vivo.
Keywords:B-Lymphoid Precursor Cells, Cell Line, Cerebellum, Down-Regulation, Embryonic Stem Cells, Gene Expression, Gene Expression Regulation, Gene Order, Gene Silencing, Gene Targeting, Multigene Family, RNA Interference, Trans-Activators, Transgenes, Animals, Mice
Source:PLoS ONE
ISSN:1932-6203
Publisher:Public Library of Science (U.S.A.)
Volume:8
Number:11
Page Range:e80312
Date:12 November 2013
Additional Information:Erratum in: PLoS ONE 8(12): 10.1371/annotation/c8b2e360-b78a-4c2f-a1a3-c53325f18211.
Official Publication:https://doi.org/10.1371/journal.pone.0080312
PubMed:View item in PubMed

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