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Antisense oligonucleotide and LNA GapmeR transfections

CR Caroline Jane Ross
AR Aviv Rom
AS Amit Spinrad
DG Dikla Gelbard-Solodkin
ND Neta Degani
IU Igor Ulitsky
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ASOs (Integrated DNA Technologies) were designed to target the conserved ATGG sites that were identified by LncLOOM in the last exon of mouse Chaserr (Additional file 1: Fig. S3A). All ASOs were modified with 2′-O-methoxy-ethyl bases. LNA gapmers (Qiagen), targeted to Chaserr introns, were used for Chaserr knockdown (see Additional file 6: Table S4 for full oligo sequences). Transfection: 2 × 105 Neuro2A cells were seeded in a six-well plate and transfected by using Lipofectamine 3000 (Life Technologies, L3000-008) following the manufacturer’s protocol with a mix of LNA1–4 or with ASO1, ASO2, ASO3, or a mix of either ASO1 and ASO3 or ASO1–3 to a final concentration of 25 nM. Endpoints for all experiments were at 48 h post transfection, after which the cells were collected with TRIZOL for RNA extraction and assessment by RT-qPCR analysis.

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