Abstract
The assessment of non-coding RNAs (ncRNAs) functions highly relies on loss of function studies. However, due to their exclusive or partial nuclear localization, many small and long ncRNAs are not efficiently silenced by RNA interference. Antisense LNA GapmeRs constitute a good alternative to RNAi. They allow an effective knockdown of ncRNAs with sizes greater than 80 nucleotides, regardless of their cellular localization. This chapter focuses on the silencing of two different nuclear ncRNAs (ANRIL and SATIII RNAs) in mammalian cells using antisense LNA GapmeRs with two different transfection methods: calcium phosphate-mediated transfection and LipofectamineTM 2000.
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Acknowledgments
This work was supported partly by the french PIA project “Lorraine Université d’Excellence” (ANR-15IDEX-04-LUE) and the RHU program FIGHT-HF (ANR-15-RHU-004). The CNRS and UL are also thanked for fundings.
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Alfeghaly, C., Aigueperse, C., Maenner, S., Behm-Ansmant, I. (2021). Non-Coding RNA Silencing in Mammalian Cells by Antisense LNA GapmeRs Transfection. In: Rederstorff, M. (eds) Small Non-Coding RNAs. Methods in Molecular Biology, vol 2300. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1386-3_4
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DOI: https://doi.org/10.1007/978-1-0716-1386-3_4
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