Abstract
Long noncoding RNAs (lncRNAs) are a functional and structural diverse class of cellular transcripts that comprise the largest fraction of the human transcriptome. However, detailed functional analysis lags behind their rapid discovery. This might be partially due to the lack of loss-of-function approaches that efficiently reduce the expression of these transcripts. Here, I describe a method that allows a specific and efficient targeting of the highly abundant lncRNA MALAT1 in human (lung) cancer cells. The method relies on the site-specific integration of RNA-destabilizing elements mediated by Zinc Finger Nucleases (ZFNs).
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References
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Acknowledgement
The author wishes to acknowledge the support of his colleagues at the German Cancer Research Center (DKFZ) Heidelberg who helped to establish this method and to set up the protocol. A special thanks goes to Matthias Groß and Dr. Monika Hämmerle for critical reading of the manuscript. T.G. is supported by an Odyssey Postdoctoral Fellowship sponsored by the Odyssey Program and the CFP Foundation at The University of Texas MD Anderson Cancer Center.
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Gutschner, T. (2015). Silencing Long Noncoding RNAs with Genome-Editing Tools. In: Pruett-Miller, S. (eds) Chromosomal Mutagenesis. Methods in Molecular Biology, vol 1239. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1862-1_13
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DOI: https://doi.org/10.1007/978-1-4939-1862-1_13
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