蛋白用于三元复合基因载体

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蛋白用于三元复合基因载体. 宋玉华 2012207474 化学工程. 基因治疗. 蛋白三元复合基因载体. 非病毒载体. 基因治疗. 治疗性蛋白. 基因物质. 特定细胞. 裸 DNA 带负电, 很难单独进入细胞,而且容易被血清中的核酸酶降解. 寻找. 载 体. 免疫原性、插入诱变. 病毒载体. 低毒、低免疫反应,外源基因随机整合 率低且携带基因大小类型不受限制. 非病毒载体. 聚阳离子. 脂质体. 阳离子 脂质体或聚阳离子 与 DNA 组装形成的 二元 复合物带有过量正电荷 , 易与血清中带负电的蛋白质非特异性结合. 不. 足. - PowerPoint PPT Presentation

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32gHSA
HSAHSADNA
HSADNA
S. Simoes et al. / Biochimica et Biophysica Acta 1463 (2000) 459~469

Transfection by HSA-lipoplexes in vivo. HSA-lipoplexes, plain lipoplexes (both at a charge ratio (+/3) of 2/1 and containing 100 Wg pCMVluc), or naked DNA were injected into mice via the tail vein in a volume of 200 Wl. Luciferase gene expression in the lungs and spleen was measured 8 h following injection, and is given as pg luciferase/mg organ. Each bar represents a dierent animal. Note that luciferase expression by naked DNA was too low to be apparent at this scale.
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Human serum albuminpolyethylenimine nanoparticles for gene delivery. Journal of Controlled Release

Effect of different N/P ratios of HSAPEIDNA nanoparticles (hatched bars) and PEIDNA complexes (black bars) on the viability of 293 cells. The MTT dye reduction assay was performed 48 h after incubation of the cells with the same amount of the preparations as in the transfection trials.
N/PHSA/PEI/DNAPEI/DNA80%
Human serum albuminpolyethylenimine nanoparticles for gene delivery. Journal of Controlled Release
HSA

J Gene Med 2005; 7: 15551564.
HSA enhances PEI-mediated transfection. 9HTEo-(A) and A549 (B) cells were plated onto 24-well plates at 100 000and 50 000 per well, respectively, to obtain 90100% confluency after 2 days of culture. Cells were incubated with 1 g of pCLuc complexed with PEI (10N/P) with HSA over a range of albumin amounts (expressed in g on the x-axis) in a total volume of 1 ml. Twenty-four hours after transfection, cells were lysed and cell lysates were tested for luciferase activity, expressed as relative light units (RLU) per g of protein content. Values represent the mean (SEM) of three experiments conducted in duplicate



Biomacromolecules 2011, 12, 10061014
BSA/PDMA/DNA50nm
BSA-PDMABSABSA-PDMA
BSA

N/P
N/P

PVPPDMAEMAPPDN/P3PPDDNAPPD/DNAPVPDNADNADNA
B. Zhang et al. / Journal of Biomaterials Science 0 (2012) 116

BSA/PPD/DNAHepG2BSA

Cytotoxicity and transfection efficiency in vitro. (a) Cell viabilities of HepG2 cells treated by BSA/PPD-1/pDNAcomplexes (the N/P ratio of the PPD-1/pDNA binary complexes is 15) with the increase of BSA concentrations determined by MTT assay. Experiments were performed in triplicate; values represent the relative viability compared to untreated cells as means SEM of one representative experiment (n = 3). (b) The transfection efficiencies of BSA/PPD-1/pDNA complexes (the N/P ratio of the PPD-1/pDNA binary complexes is 15) with the increase of BSA concentrations in HepG2 cells determined by flow cytometry. Experiments were performed in triplicate; values represent the EGFP-positive cells as means SEM of one representative experiment (n = 3). (c) Fluorescence images of HepG2 cells. (c1, c2) PPD-1/pDNA binary complexes at a N/P ratio of 15 and BSA/PPD-1/pDNA ternary complexes (the concentration of BSA is 7.5 g/ml), respectively.
10%HepG2BSA/PPD/DNAPEI
BSADNA

PEI/DNANIH/3T3/DNA
Journal of Nanoscience and Nanotechnology, 2010, 10(5): 3170-3174.

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