The lncRNA VELUCT strongly regulates viability of lung cancer cells despite its extremely low abundance
Little is known about the function of most non-coding RNAs (ncRNAs). The majority of long ncRNAs (lncRNAs) is expressed at very low levels and it is a matter of intense debate whether these can be of functional relevance. Here, we identified lncRNAs regulating the viability of lung cancer cells in a...
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| Hauptverfasser: | , , , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
04 February 2017
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| In: |
Nucleic acids research
Year: 2017, Jahrgang: 45, Heft: 9, Pages: 5458-5469 |
| ISSN: | 1362-4962 |
| DOI: | 10.1093/nar/gkx076 |
| Online-Zugang: | Verlag, Volltext: http://dx.doi.org/10.1093/nar/gkx076 Verlag, Volltext: https://academic.oup.com/nar/article/45/9/5458/2970150 
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| Verfasserangaben: | Jana Seiler, Marco Breinig, Maïwen Caudron-Herger, Maria Polycarpou-Schwarz, Michael Boutros and Sven Diederichs |
MARC
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| 520 | |a Little is known about the function of most non-coding RNAs (ncRNAs). The majority of long ncRNAs (lncRNAs) is expressed at very low levels and it is a matter of intense debate whether these can be of functional relevance. Here, we identified lncRNAs regulating the viability of lung cancer cells in a highthroughput RNA interference screen. Based on our previous expression profiling, we designed an siRNA library targeting 638 lncRNAs upregulated in human cancer. In a functional siRNA screen analyzing the viability of lung cancer cells, the most prominent hit was a novel lncRNA which we called Viability Enhancing LUng Cancer Transcript (VELUCT). In silico analyses confirmed the non-coding properties of the transcript. Surprisingly, VELUCT was below the detection limit in total RNA from NCI-H460 cells by RT-qPCR as well as RNA-Seq, but was robustly detected in the chromatin-associated RNA fraction. It is an extremely low abundant lncRNA with an RNA copy number of less than one copy per cell. Blocking transcription with actinomycin D revealed that VELUCT RNA was highly unstable which may partially explain its low steady-state concentration. Despite its extremely low abundance, loss-of-function of VELUCT with three independent experimental approaches in three different lung cancer cell lines led to a significant reduction of cell viability: Next to four individual siRNAs, also two complex siPOOLs as well as two antisense oligonucleotides confirmed the strong and specific phenotype. | ||
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