Journal: Biomacromolecules

Article Title: Highlighting the Role of Polymer Length, Carbohydrate Size, and Nucleic Acid Type in Potency of Glycopolycation Agents for pDNA and siRNA Delivery

doi: 10.1021/bm401026n

Figure Lengend Snippet: Figure 5. Influence of siRNA concentration and N/P ratio on polyplex-induced target gene (luciferase) down-regulation in U-87_luc2 glioblastoma cells. Notation is as follows: hashed bar (lighter color) indicates treatment with polyplexes containing siRNA and the solid bar (darker color) indicates treatment with polyplexes containing siCon. The numbers above the bars indicate percentage of gene down-regulation. The notation Tr4(77) 50/100 indicates that polyplexes were formulated with siRNA and Tr4(77) at an N/P ratio of 50 and with 100 nM siRNA concentration. “siLuc2” indicates antiluciferase siRNA; “siCon” indicates scrambled siRNA negative control. (A) Knockdown efficiency response to polyplexes formed with Tr4(77) with 100 nM siRNA at differing N/P ratio; (B) Extent of knockdown response to Tr4(77) polyplexes formulated at an N/P ratio of 50 at siRNA concentrations from 1 to 100 nM; (C) Knockdown efficiency response to polyplexes formed with CD4(143) with 100 nM siRNA at differing N/P ratios; (D) Extent of knockdown response to CD4(143) formulated at an N/P ratio of 50 at siRNA concentrations from 1 to 100 nM. [Note: The “*” symbol indicates significantly different values as compared to “cells only” control (p < 0.05).]

Article Snippet: U-87 MG-luc2 (U-87_luc2) cells, human glioblastoma cells genetically engineered to constitutively express luciferase, were obtained from Caliper Life Sciences, Inc. (now Perkin Elmer, Inc., Mountain View, CA).

Techniques: Concentration Assay, Luciferase, Negative Control, Knockdown, Control

Journal: Biomacromolecules

Article Title: Highlighting the Role of Polymer Length, Carbohydrate Size, and Nucleic Acid Type in Potency of Glycopolycation Agents for pDNA and siRNA Delivery

doi: 10.1021/bm401026n

Figure Lengend Snippet: Figure 7. siRNA-mediated gene down-regulation with U87-luc2 cells as a function of polyplex type (formed with either Tr4 or CD4 polymers) and polymer degree of polymerization. (A) Luciferase gene down-regulation facilitated by polyplexes formulated with polymer Tr4 (nw = 23, 55, 77) at an N/P = 50 using 100 nM siRNA (50/100). (B) Uptake of Cy5-labeled siRNA within Tr4-containing polyplexes formulated at an N/P = 50 with 40 nM of siRNA (50/40) as measured by flow cytometry 3 h after transfection. (C) Luciferase gene down-regulation facilitated by polyplexes formulated with polymer CD4 (nw = 10, 26, 39, 143, 239) at an N/P = 50 using 100 nM siRNA (50/100). (D) Uptake of Cy5-labeled siRNA within CD4-containing polyplexes formulated at an N/P = 50 with 40 nM of siRNA (50/40) as measured by flow cytometry 3 h after transfection. In graphs A and C, the hashed bars indicate data for cells treated with siRNA-containing polyplexes, and the solid bars indicate data for cells treated with siCon-containing polyplexes (control). The numbers above the bars indicate percentage of gene down-regulation. [Note the “*” symbol indicates significantly different values as compared to the cells only sample (p < 0.05).]

Article Snippet: U-87 MG-luc2 (U-87_luc2) cells, human glioblastoma cells genetically engineered to constitutively express luciferase, were obtained from Caliper Life Sciences, Inc. (now Perkin Elmer, Inc., Mountain View, CA).

Techniques: Polymer, Luciferase, Labeling, Cytometry, Transfection, Control

Journal: Biomacromolecules

Article Title: Highlighting the Role of Polymer Length, Carbohydrate Size, and Nucleic Acid Type in Potency of Glycopolycation Agents for pDNA and siRNA Delivery

doi: 10.1021/bm401026n

Figure Lengend Snippet: Figure 8. Comparison of pDNA and siRNA delivery efficiency for luciferase expression and down-regulation, respectively, in the absence and presence of serum. (A, B) Delivery of pDNA encoding the firefly luciferase reporter gene into U87 glioblastoma cells with the carbohydrate- containing polymers and controls in (A) OptiMEM and (B) DMEM. Trehalose (Tr4) or β-cyclodextrin (CD4)-containing polyplexes were formulated with pDNA at N/P = 7, while pDNA complexes with T4 or G4 were formulated at N/P = 20 (maximum gene expression as previously published).17,22 (C, D) Delivery of antiluciferase siRNA into U87-luc2 glioblastoma cells with the carbohydrate-containing polymers and controls in (C) OptiMEM and (D) DMEM. All of the carbohydrate-containing polymers were formulated at a siRNA concentration of 100 nM and N/P = 50. [Note the “*” symbol indicates significantly different values as compared to the “cells only” sample (p < 0.05).]

Article Snippet: U-87 MG-luc2 (U-87_luc2) cells, human glioblastoma cells genetically engineered to constitutively express luciferase, were obtained from Caliper Life Sciences, Inc. (now Perkin Elmer, Inc., Mountain View, CA).

Techniques: Comparison, Luciferase, Expressing, Gene Expression, Concentration Assay