Journal: Frontiers in Neuroscience

Article Title: Improving the Delivery of SOD1 Antisense Oligonucleotides to Motor Neurons Using Calcium Phosphate-Lipid Nanoparticles

doi: 10.3389/fnins.2017.00476

Figure Lengend Snippet: Physical properties of SOD1-ASO-loaded CaP-lipid NPs with a Ca/P ratio 100:1. Schematic representation of SOD1-ASO loaded CaP-lipid NP formulation (A) , particle size (d.nm) of SOD1-ASO loaded (black) and non-loaded (red) CaP-lipid NPs determined by DLS (B) , zeta potential of SOD1-ASO CaP-lipid NP (C) , transmission electron micrograph (TEM) of SOD1-ASO CaP-lipid NPs visualized by negative staining (D) , pH-sensitivity as indicated by change in number size (d.nm) of CaP-lipid NPs in PBS at pH 7.4, 6.5, and 5.0 following incubation at 37°C for up to 4 h (E) . ** P < 0.01.

Article Snippet: SOD1 antisense oligonucleotide (target sequence 5′-CCG TCG CCC TTC AGC ACG CA-3′) (Patent US 20120214865 A1) and a scrambled negative control oligonucleotide (target sequence 5'-GCC AGC CTA CGA CTC CGC TC-3′) were synthesized by GeneWorks.

Techniques: Formulation, Zeta Potential Analyzer, Transmission Assay, Negative Staining, Incubation

Journal: Frontiers in Neuroscience

Article Title: Improving the Delivery of SOD1 Antisense Oligonucleotides to Motor Neurons Using Calcium Phosphate-Lipid Nanoparticles

doi: 10.3389/fnins.2017.00476

Figure Lengend Snippet: Particle size, Zeta (ζ)-potential, polydispersity index, and encapsulation effiency of SOD1 antisense oligonucleotide calcium phosphate-lipid nanoparticles (SOD1-ASO CaP-lipid NPs) prepared at different CaCl 2 , and Na 2 HPO 4 concentrations.

Article Snippet: SOD1 antisense oligonucleotide (target sequence 5′-CCG TCG CCC TTC AGC ACG CA-3′) (Patent US 20120214865 A1) and a scrambled negative control oligonucleotide (target sequence 5'-GCC AGC CTA CGA CTC CGC TC-3′) were synthesized by GeneWorks.

Techniques: Encapsulation, Zeta Potential Analyzer, Concentration Assay

Journal: Frontiers in Neuroscience

Article Title: Improving the Delivery of SOD1 Antisense Oligonucleotides to Motor Neurons Using Calcium Phosphate-Lipid Nanoparticles

doi: 10.3389/fnins.2017.00476

Figure Lengend Snippet: Internalization of CaP-lipid NPs into NSC-34 cells and knockdown of SOD1 protein expression in HEK293 cells. The CaP nanoparticles contained LissRdB-DSPE (Ex = 560 nm; Em = 583 nm) in the outer lipid layer. NSC-34 cells were incubated at 37°C with LissRdB-DSPE CaP-lipid NPs diluted either 1:5 or 1:10 (v/v) for 30, 60 (A) and 90 min (B) . Images were acquired with a 63 × objective (512 × 512 pixels; physical length 246.03 μm × 246.03 μm). Change in average cell fluorescence over 90 min incubation period (C) . Red dotted line represents auto fluorescence of NSC-34 cells for comparison. *** P < 0.001 and # P < 0.05 signifies significant difference compared to respective 30 min data. HEK293 cells were treated with SOD1-ASO CaP-lipid NPs for 72 h at 37°C (Lane 3). Control treatments included non-loaded CaP-lipid NPs (Lane 2) and non-loaded CaP-lipid NPs with SOD1-ASO free in solution (Lane 1). Controls using Lipofectamine 2000 Transfection Reagent with SOD1-ASO or scrambled negative SOD1 oligonucleotide are not shown here. The change in SOD1 protein expression (16 kDa) was visualized sing SuperSignal West Pico Chemiluminescent Substrate and exposed using an Amersham 600RGB Imager (D) .

Article Snippet: SOD1 antisense oligonucleotide (target sequence 5′-CCG TCG CCC TTC AGC ACG CA-3′) (Patent US 20120214865 A1) and a scrambled negative control oligonucleotide (target sequence 5'-GCC AGC CTA CGA CTC CGC TC-3′) were synthesized by GeneWorks.

Techniques: Knockdown, Expressing, Incubation, Fluorescence, Comparison, Control, Transfection