Non-Coding RNA Silencing in Mammalian Cells by Antisense LNA GapmeRs Transfection.
Fiche publication
Date publication
janvier 2021
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Dr BEHM-ANSMANT Isabelle
Tous les auteurs :
Alfeghaly C, Aigueperse C, Maenner S, Behm-Ansmant I
Lien Pubmed
Résumé
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 Lipofectamine 2000.
Mots clés
ANRIL, Antisense LNA GapmeRs, Calcium phosphate transfection, LipofectamineTM 2000, SATIII, lncRNA silencing
Référence
Methods Mol Biol. 2021 ;2300:31-37