Big Chemical Encyclopedia

Chemical substances, components, reactions, process design ...

Articles Figures Tables About

Lipoplex transfection

Fig. 3. In vitro transfection activity by AF-lipoplexes. Transfection of FlepG2 cells by plain (nontargeted) and AF-lipoplexes as a function of fhe amounf of the ligand. The data represent the mean SD of three wells and are representative of three independent experiments (10)... Fig. 3. In vitro transfection activity by AF-lipoplexes. Transfection of FlepG2 cells by plain (nontargeted) and AF-lipoplexes as a function of fhe amounf of the ligand. The data represent the mean SD of three wells and are representative of three independent experiments (10)...
Sakaguchi N, Kojima C, Harada A, Koiwai K, Kono K (2008) The correlation between fusion capability and transfection activity in hybrid complexes of lipoplexes and pH-sensitive liposomes. Biomaterials 29 4029 1036... [Pg.27]

The administration of SPLP results in reporter gene expression at the tumor site (Fig. 7B). Injection of free plasmid or lipoplexes resulted in no detectable gene expression at the tumor site. However, transfection was observed in the limg, liver, and spleen. SPLP, on the other hand, did not show detectable levels of gene expression in these organs. [Pg.143]

Despite the fact that many different cationic lipids have been synthesized and tested for transfection (25 34), relatively few systematic structure activity TE-relationship studies have been performed (35 39). As a result, no general relationship between chemical structure and TE could be drawn from these studies. One reason for this is that the chemical structure of a cationic lipid is not directly responsible for TE. TE rather depends on the biophysical characteristics of the cationic lipid aggregate (e.g., liposomes and lipoplexes), which, for its part, is dependent on the chemical structure of the lipids. In a previous study with analogs of the transfection lipid A-[l-(2,3-dioleoyloxy) propyl]-A,A,A-trimethylammoniumchloride (DOTAP) (40) which differ in their nonpolar hydrocarbon chains, it could be shown that the TE strongly depended on the biophysical properties of the resulting liposomes and lipoplexes (35). Minimal alterations of biophysical properties by using lipids with different hydrocarbon chains or by mixing the lipid with different neutral helper lipids could completely allow or prevent transfection. [Pg.254]

As indicated in Figure 1, the process of lipofection can be divided into independent steps (i) preparation of a lipofection reagent, (ii) formation of lipoplexes, and (iii) the transfection itself. [Pg.254]

Figure 1 The principles and variant parameters of lipofection. (i) Preparation of a lipofection reagent cationic liposomes were prepared from cationic lipids and helper (if required), (ii) Formation of positively charged lipoplexes by addition of DNA (e.g., reporter plasmid carrying the firefly luciferase gene) to the cationic liposomes, (iii) Transfection (lipofection) by incubation cells with the preformed lipoplexes. The efficiency of gene transfer (lipofection efficiency) can be determined from reporter gene amount or activity (e.g., luciferase activity). Most of the steps of a lipofection experiment can be varied and optimized (grey spots). Figure 1 The principles and variant parameters of lipofection. (i) Preparation of a lipofection reagent cationic liposomes were prepared from cationic lipids and helper (if required), (ii) Formation of positively charged lipoplexes by addition of DNA (e.g., reporter plasmid carrying the firefly luciferase gene) to the cationic liposomes, (iii) Transfection (lipofection) by incubation cells with the preformed lipoplexes. The efficiency of gene transfer (lipofection efficiency) can be determined from reporter gene amount or activity (e.g., luciferase activity). Most of the steps of a lipofection experiment can be varied and optimized (grey spots).
COS-7 or CHO cells (for initial transfection screening) or cells of therapeutic interest (e.g., dendritic cells and various cancer cells) at a confluence of 50%, grown in 96-well culture plates, were placed into the robot by the robotic conveyor. In a fully automated process, the robot removes the lid from the cell culture microtiter plate, dispenses lipoplexes into the wells (triplicates), replaces the lid and returns the plate to the incubator. After four hours, the cells are automatically retrieved, the cell monolayers are carefully washed using a special drop mode of the integrated plate washer, fresh medium is added, and the cells are incubated for further 42 hours before harvesting. [Pg.261]

The previous screening experiments were performed with lipoplexes containing equimolar amounts of the helper lipid DOPE. Here, the influence of different ratios of the helper lipids DOPE and Choi on TE of KL-1-14 were tested. The transfection behavior of KL-1-14 without any helper lipid was tested as well. [Pg.265]

TE of KL-1-14 without helper lipids was very low and reached only about twice the TE, which was found for the standard lipid DOTAP. Independent of the amount of DOPE incorporated in the lipoplexes (ratio of DOPE/KL-1-14 0.3, 0.5, 0.6, 0.7, 0.8, 0.8, 1.0, and 1.2), transfection behaviors (maximum transfection efficiencies and transfection profiles) of all mixtures were similar and comparable to the profile of KL-1-14/DOPE (1 1) as shown in Figure 2 (individual data for all mixtures are not shown). [Pg.265]

Figure 6 Lipofection results (lipofection profiles) of lipoplexes from the R-configu-rated cationic lipids KL-1-1 to KL-1-17 (Table 1) in a mixture with equimolar amounts of l,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (DOPE) (counterion chloride) and the pCMVluc-plasmid. Each bar represents the mean ( S.D.) of three wells of a 96-well microtiter plate. T-axis (left) represents the transfection efficiencies expressed in relative light units (RLU) (lu/pg protein). X-axis (right) represents the viability of the cells compared to nontreated control cells. F-axis represents the different cationic lipid/plasmid DNA-charge ratios from 1 to 15. Figure 6 Lipofection results (lipofection profiles) of lipoplexes from the R-configu-rated cationic lipids KL-1-1 to KL-1-17 (Table 1) in a mixture with equimolar amounts of l,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (DOPE) (counterion chloride) and the pCMVluc-plasmid. Each bar represents the mean ( S.D.) of three wells of a 96-well microtiter plate. T-axis (left) represents the transfection efficiencies expressed in relative light units (RLU) (lu/pg protein). X-axis (right) represents the viability of the cells compared to nontreated control cells. F-axis represents the different cationic lipid/plasmid DNA-charge ratios from 1 to 15.
Transfection efficiencies of the KL-1-14 lipoplexes were compared to the TE achieved with the standard transfection lipid DOTAP. Results were given in RLU (lu/pg protein) and, for easier comparison, standardized on the lipofection efficiency of DOTAP-lipoplexes, which was set to 100% Compared to the respective DOTAP-value. [Pg.268]

Dass CR. Lipoplex-mediated delivery of nucleic acids factors affecting in vivo transfection. J Mol Med 2004 82(9) 579-591. [Pg.270]

Numerous studies reported the in vitro interaction of lipoplexes with serum (14). The parameters checked, such as maintained lipid-DNA interactions, aggregate formation, zeta potential changes, or DNA morphological changes, did not allow for consistent prediction of in vitro transfection efficiency of lipoplexes in serum (15). However, it is obviously an indication of poor delivery of DNA at the target site. [Pg.275]

In addition to extracellular degradation in tissues, endosomal acidification might also trigger PEG-lipid cleavage. We showed that despite the presence of the PEG, which slightly reduces lipoplex internalization into the cells, DNA transfection level almost reaches the level of the cationic lipo-plex (31). Cholesterol PEG incorporation into lipoplexes not only reduces lipoplex internalization, but also inhibits the transfection efficiency. [Pg.281]

Faneca H, Simoes S, Pedroso de Lima MC. Association of albumin or protamine to lipoplexes enhancement of transfection and resistance to serum. J Gene Med 2004 6 681. [Pg.290]

Tandia B, et al. Lipid mixing between lipoplexes and plasma lipoproteins is a major barrier for intravenous transfection mediated by cationic lipids. J Biol Chem 2005 280 12255. [Pg.290]

Rejman J, et al. Characterization and transfection properties of lipoplexes stabilized with novel exchangeable polyethylene glycol-lipid conjugates. Biochim Biophys Acta 2004 1660 41. [Pg.292]

Inclusion of an NLS consensus peptide into a lipoplex renders nondividing cells susceptible to gene transfection. There is dramatic improvement... [Pg.306]

Liposomes can be modified in numerous fashions by the addition of peptide sequences. The benefit of this is that the inclusion of the peptide allows the lipoplex to be optimized for its task such as the targeting a specific cell type with a specific receptor ligand sequence or the transfection of nondividing cells, with an NLS. [Pg.307]

Simoes S, Slepushkin V, Pretzer E, et al. Transfection of human macrophages by lipoplexes via the combined use of transferrin and pH-sensitive peptides. J Leukoc Biol 1999 65(2) 270-279. [Pg.380]

Mortimer et al., 1999). In contrast, the level of reporter gene expression was enhanced when cells were exposed to lipoplexes during or just before mitosis (Brunner et al., 2000). Finally, the relatively low rate of cell proliferation of primary cultures of ciliated human airway epithelia was found to be one of the determinants for their low transfectability by lipoplex (Fasbender et al., 1997). These results, however, do not preclude the possibility that DNA can be translocated through nuclear pore complex by a NLS-independent mechanism. [Pg.199]

Audouy S, Molema G, de Leij L, Hoekstra D (2000) Serum as a modulator of lipoplex-mediated gene transfection dependence of amphiphile, cell type and complex stability. J Gene Med 2(6) 465 176... [Pg.13]


See other pages where Lipoplex transfection is mentioned: [Pg.1044]    [Pg.1044]    [Pg.137]    [Pg.138]    [Pg.246]    [Pg.254]    [Pg.255]    [Pg.256]    [Pg.257]    [Pg.261]    [Pg.265]    [Pg.269]    [Pg.274]    [Pg.274]    [Pg.276]    [Pg.282]    [Pg.282]    [Pg.351]    [Pg.192]    [Pg.198]    [Pg.227]    [Pg.290]    [Pg.322]    [Pg.463]    [Pg.463]    [Pg.7]    [Pg.8]    [Pg.8]    [Pg.18]    [Pg.24]   
See also in sourсe #XX -- [ Pg.430 , Pg.431 ]




SEARCH



Lipoplex

Lipoplexes

Transfectants

© 2024 chempedia.info