nusinersen  (Biogen Inc)

Journal: bioRxiv

Article Title: Sustained Relief of Chronic Pain via a Nav1.7-Targeting ASO–siRNA Conjugate

doi: 10.64898/2026.03.27.714734

Figure Lengend Snippet: (A, B) Primary screening in SK-N-AS cells. Nav1.7 mRNA levels were quantified by RT-qPCR 48 h after transfection with RNAiMAX. ASOs were tested at 10 and 100 nM and ranked by knockdown efficiency at 100 nM ( A) . siRNAs were tested at 0.1 and 10 nM and ranked by knockdown efficiency at 10 nM ( B ). ( C, D) Immunostimulatory potential of N02A114 in human PBMCs treated with 0.078, 0.31, or 1.25 μM ASO. IL-6 ( C ) and TNF-α ( D ) levels were measured after 24 h by ELISA. Nusinersen and tandem CpG oligonucleotides served as negative (NC) and positive (PC) controls, respectively. ( E, F) Cytotoxicity assessment by caspase-3/7 activity in Hepa1-6 ( E ) and HeLa ( F ) cells 24 h after transfection with N02A114 (6.25, 12.5, or 25 nM). Nusinersen served as NC and the hepatotoxic ASO 558807 as PC. ( G) Seed-mediated off-target activity of 12 siRNAs measured in HeLa cells using a dual-luciferase reporter assay at 0.39-25 nM. ( H, I) Dose-response curves for N02A114 ( H ) and N02S154 ( I ) in SK-N-AS and rat DRG cells determined by RT-qPCR. n = 3. Data represent one of three independent experiments.

Article Snippet: Nusinersen (Cat. No. HY-112980) was purchased from MedChemExpress.

Techniques: Quantitative RT-PCR, Transfection, Knockdown, Enzyme-linked Immunosorbent Assay, Activity Assay, Luciferase, Reporter Assay

Journal: bioRxiv

Article Title: Sustained Relief of Chronic Pain via a Nav1.7-Targeting ASO–siRNA Conjugate

doi: 10.64898/2026.03.27.714734

Figure Lengend Snippet: (A) Screening of ASOs in rat DRG cells at a concentration of 1,000 nM. ASOs were delivered to DRG cells by free uptake, and Nav1.7 mRNA levels were measured 72 h after treatment by RT-qPCR. ASOs were ranked according to their knockdown activity. n = 3 experimental replicates. (B-M) Dose-response curves for 12 siRNAs in rat DRG cells. Cells were treated via free uptake and Nav1.7 mRNA levels were quantified by RT-qPCR. n = 3 experimental replicates. (N) Immunostimulatory assessment of selected ASOs in human peripheral blood mononuclear cells (PBMCs). Eighteen ASOs were incubated with PBMCs at 0.31 or 1.25 μM for 24 h, and IL-6 levels were measured using a sandwich ELISA. Nusinersen and tandem CpG oligodeoxynucleotides served as the negative control (NC) and positive control (PC), respectively. n = 3 experimental replicates. (O, P) Caspase 3/7 activity in Hepa1-6 ( O ) and Hela ( P ) cells was measured at 24 h after transfection of 17 selected ASOs at 25 nM. A reported sequence (558807) served as a PC and Nusinersen served as a NC. n = 3 experimental replicates. Data are presented as mean ± SEM.

Article Snippet: Nusinersen (Cat. No. HY-112980) was purchased from MedChemExpress.

Techniques: Concentration Assay, Quantitative RT-PCR, Knockdown, Activity Assay, Incubation, Sandwich ELISA, Negative Control, Positive Control, Transfection, Sequencing