Table 1.
Vectors used during transfection studies and gene delivery comparison
| Vector | MOI | D9 dosage, mg | Vector size, nm | No. of vectors per well | pDNA per vector, ng | Total pDNA, ng | Gene delivery (GD), luminescence per μg of protein | Delivery efficiency, GD per total ng of pDNA | GFP-positive cells | Delivery efficiency × % of GFP cells | Viable cells | Delivery efficiency × % of GFP cells × % of viable cells (TGDP) |
| S1: D9 hybrid | 1:1 | 0 | ∼2,000 | 30,000 | 2.7 × 10−7* | 0.008 | 8 | 970 | 8% | 78 | 99% | 77 |
| 0.1 | 11 | 1,400 | ||||||||||
| 0.25 | 15 | 1,880 | ||||||||||
| 0.4 | 19 | 2,370 | 10% | 240 | 100% | 240 | ||||||
| 0.5 | 19 | 2,410 | ||||||||||
| 1 | 22 | 2,740 | ||||||||||
| 10:1 | 0 | ∼2,000 | 300,000 | 2.7 × 10−7* | 0.08 | 110 | 1,360 | 32% | 430 | 82% | 350 | |
| 0.1 | 180 | 2,250 | ||||||||||
| 0.25 | 180 | 2,280 | ||||||||||
| 0.4 | 220 | 2,730 | 27% | 730 | 88% | 650 | ||||||
| 0.5 | 170 | 2,090 | ||||||||||
| 1 | 130 | 1,630 | ||||||||||
| 100:1 | 0 | ∼2,000 | 3,000,000 | 2.7 × 10−7* | 0.8 | 1,190 | 1,480 | 79% | 1,170 | 55% | 640 | |
| 0.1 | 2,350 | 2,930 | ||||||||||
| 0.25 | 2,460 | 3,070 | ||||||||||
| 0.4 | 2,360 | 2,940 | 67% | 1,970 | 58% | 1,140 | ||||||
| 0.5 | 1,820 | 2,260 | ||||||||||
| 1 | 1,580 | 1,960 | ||||||||||
| D9 polyplex | N/A | 0.018 | 150 | 8.3 × 109† | 7.2 × 10−8 | 600 | 30 | 0.051 | 7% | 0.003 | 94% | 0.003 |
| D9-Man polyplex | N/A | 0.018 | 150 | 8.3 × 109† | 7.2 × 10−8 | 600 | 40 | 0.067 | 23% | 0.015 | 95% | 0.015 |
| FuGENE 6 (Promega) | N/A | N/A | N/A | N/A | N/A | 200 | 44 | 0.219 | 31% | 0.068 | 100% | 0.068 |
| FuGENE HD (Promega) | N/A | N/A | N/A | N/A | N/A | 200 | 100 | 0.521 | 55% | 0.285 | 100% | 0.285 |
| ViaFECT (Promega) | N/A | N/A | N/A | N/A | N/A | 100 | 22 | 0.225 | 16% | 0.035 | 100% | 0.035 |
| OmniFect (TransOMIC Technologies) | N/A | N/A | N/A | N/A | N/A | 100 | 38 | 0.375 | 20% | 0.075 | 100% | 0.075 |
| Xfect (Clontech Laboratories) | N/A | N/A | N/A | N/A | N/A | 500 | 33 | 0.067 | 57% | 0.038 | 78% | 0.030 |
| JET-PEI (Polyplus transfection) | N/A | N/A | N/A | N/A | N/A | 250 | 130 | 0.511 | 30% | 0.152 | 85% | 0.129 |
| GeneJuice (EMD Millipore) | N/A | N/A | N/A | N/A | N/A | 100 | 3 | 0.027 | 1% | 0.000 | 100% | 0.000 |
N/A, not applicable.
*
Amount of bacterially maintained pDNA was estimated using a pBR322 origin, which is regulated at 20–40 copies per bacterium, and the molecular weight of the plasmid [estimated as plasmid base pairs × 650 (daltons per base pair)].
†
Polyplex number was calculated by dividing the total mass of D9 added per well at a 30:1 polymer-to-pDNA ratio by an estimated mass of one polyplex. Individual polyplex mass was estimated using the measured effective diameter to calculate volume and a density of 1.22 g/mL (31).