全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...
科学通报  2014 

非病毒载体介导的siRNA细胞内导入机理研究

DOI: 10.1360/972013-639, PP. 188-194

Keywords: 超支化聚酰胺-胺,siRNA输送,基因沉默,网格蛋白介导的内吞作用,窖蛋白介导的内吞,内涵体逃逸

Full-Text   Cite this paper   Add to My Lib

Abstract:

比较了两类非病毒载体,包括阳离子聚合物载体(HPAMAM-PHE45,PEI)和阳离子脂质载体(DOTAP)运载siRNA进入细胞的动态过程.DOTAP/siRNA能够很快地被内吞进入内涵体途径,6h内就能将绝大多数的siRNA从内涵体中释放出来.而HPAMAM-PHE45,PEI/siRNA复合物在与细胞接触0.5h后首先附着在细胞膜表面,逐渐地部分复合物也经由内涵体途径将siRNA从内涵体中释放出来进入细胞质.但也有一部分复合物可能从窖蛋白介导的内吞途径进入细胞.实验结果显示,细胞摄取阳离子脂质/siRNA主要通过网格蛋白介导的内吞实现,而阳离子聚合物/siRNA主要通过网格蛋白介导的内吞和窖蛋白介导的内吞共同完成.这两种不同的内吞方式可能对非病毒载体的不同转染机理及优化理论有较大影响.深入研究其中的关键因素可以为非病毒载体运载siRNA内吞和胞内途径的机理提供线索.

References

[1]  2 Friend D S, Papahadjopoulos D, Debs R J. Endocytosis and intracellular processing accompanying transfection mediated by cationic liposomes. Biochim Biophys Acta, 1996, 1278: 41-50
[2]  3 Zuhorn I S, Kalicharan R, Hoekstra D. Lipoplex-mediated transfection of mammalian cells occurs through the cholesterol-dependent clathrin-mediated pathway of endocytosis. J Biol Chem, 2002, 277: 18021-18028
[3]  4 Farhood H, Serbina N, Huang L. The role of dioleoylphosphatidyle-thanolamine in cationic liposome mediated gene transfer. Biochim Biophys Acta, 1995, 1235: 289-295
[4]  6 Smisterova J, Wagenaar A, Stuart M C, et al. Molecular shape of the cationic lipid controls the structure of cationic lipid/dioleylphospha-tidylethanolamine-DNA complexes and the efficiency of gene delivery. J Biol Chem, 2001, 276: 47615-47622
[5]  8 Haensler J, Szoka F C Jr. Polyamidoamine cascade polymers mediate efficient transfection of cells in culture. Bioconjugate Chem, 1993, 4: 372-379
[6]  9 Boussif O, Lezoualc'h F, Zanta M A, et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: Polyethylenimine. Proc Natl Acad Sci USA, 1995, 92: 7297-7301
[7]  10 Behr J-P. The proton sponge: A trick to enter cells the viruses did not exploit. Chimia, 1997, 51: 34-36
[8]  13 Hansen S H, Sandvig K, van Deurs B. Clathrin and HA2 adaptors: Effects of potassium depletion, hypertonic medium, and cytosol acidification. J Cell Biol, 1993, 121: 61-72
[9]  14 Rejman J, Bragonzi A, Conese M. Role of Clathrin-and Caveolae-mediated endocytosis in gene transfer mediated by lipo-and polyplexes. Mol Ther, 2005, 12: 468-474
[10]  15 Lin P J, Tam Y Y, Hafez I, et al. Influence of cationic lipid composition on uptake and intracellular processing of lipid nanoparticle formulations of siRNA. Nanomed-Nanotechnol, 2013, 9: 233-246
[11]  16 Nel A E, M?dler L, Velegol D, et al. Understanding biophysicochemical interactions at the nano-biointerface. Nat Mater, 2009, 8: 543-557
[12]  1 Bumcrot D, Manoharan M, Koteliansky V, et al. RNAi therapeutics: A potential new class of pharmaceutical drugs. Nat Chem Biol, 2006, 2: 711-719
[13]  5 Koltover I, Salditt T, Radler J O, et al. An inverted hexagonal phase of cationic liposome-DNA complexes related to DNA release and delivery. Science, 1998, 281: 78-81
[14]  7 Xu Y, Szoka F C Jr. Mechanism of DNA release from cationic liposome/DNA complexes used in cell transfection. Biochemistry, 1996, 35: 5616-5623
[15]  11 Wang X, He Y J, Wu J, et al. Synthesis and evaluation of phenylalanine-modified hyperbranched poly(amido amine)s as promising gene carriers. Biomacromolecules, 2010, 11: 245-251
[16]  12 Hillaireau H, Couvreur P. Nanocarriers' entry into the cell: Relevance to drug delivery. Cell Mol Life Sci, 2009, 66: 2873-2896

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133