DNA initiates polymorphic structural transitions in lecithin

Tarahovsky, Yury S.; Khusainova, R. S.; Gorelov, Alexander; Nicolaeva, T.I.; Deev, A. A.; Dawson, A.K.; Ivanitsky, G. R.

doi:10.1016/0014-5793(96)00643-6

Cited by 45 publications

(56 citation statements)

References 25 publications

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“…It has been shown that such an organization can induce the formation of a very clustered form of DNA (40). Studies on model membrane systems showed that DNA-lipid complexes could be formed under different conditions bringing DNA interfacial accumulation (41,42). The local interfacial high concentration of DNA induced an electrostatic segregation of the cationic lipids and a compaction of the outer layer (16).…”

Section: Discussionmentioning

confidence: 99%

Direct visualization at the single-cell level of electrically mediated gene delivery

Golzio

Teissié

Rols

2002

Proc. Natl. Acad. Sci. U.S.A.

373

290

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Electropermeabilization is one of the nonviral methods successfully used to transfer genes into living cells in vitro and in vivo. Although this approach shows promise in the field of gene therapy, very little is known about the basic processes supporting DNA transfer. The present investigation studies this process at the single-cell level by using digitized fluorescence microscopy. Permeabilization is a prerequisite for gene transfer. Its assay by propidium-iodide (PI) penetration shows that it occurs at the sides of the cell membrane facing the two electrodes, whereas fluorescently labeled plasmids only interact with the electropermeabilized side of the cell facing the cathode. The plasmid interaction with the electropermeabilized part of the cell surface results in the formation of localized aggregates. These membrane-associated spots are formed only when pulses with a longer duration than a critical value are applied. These complexes are formed within 1 s after the pulses and cannot be destroyed by pulses of reversed polarities. They remain at the membrane level up to 10 min after pulsing. Although freely accessible to DNA dye (TOTO-1) 1 min after the pulses, they are fully protected when the addition takes place 10 min after. They diffuse in the cytoplasm 30 min after pulses and are present around the nucleus 24 h later.

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Section: Discussionmentioning

confidence: 99%

Direct visualization at the single-cell level of electrically mediated gene delivery

Golzio

Teissié

Rols

2002

Proc. Natl. Acad. Sci. U.S.A.

373

290

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“…Electron freeze fracture micrographs of aggregates of DNA formed with natural egg yolk phosphatidylcholine (EYPC) and synthetic saturated DPPC in presence of Ca 2+ cations suggested structures with long-range organization. The honeycomb-like and spaghetti-like structures were observed in EYPC+DNA+Ca 2+ aggregates (Tarahovsky et al, 1996), while structures formed in presence of DPPC were identified either as sandwich or spaghetti-like, depending on the DNA base:DPPC molar ratio . Small-angle X-ray diffractograms show two possible structural organizations of DPPC+DNA+divalent cation aggregates: We identified a sandwich structure in the aggregate DPPC:DNA:Mg 2+ =1:1:5 mol/base/mol in the temperature range 20-60°C, with a lipid bilayer repeat distance d~8.0-7.6 nm and interhelical DNA -DNA distance d DNA~6 .4-6.0 nm (Uhríková et al, 2001).…”

Section: Introductionmentioning

confidence: 97%

The structural diversity of DNA–neutral phospholipids–divalent metal cations aggregates: a small-angle synchrotron X-ray diffraction study

Uhrı́ková

Lengyel

Hanulová³

et al. 2006

Eur Biophys J

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“…However, complexes composed entirely of natural, zwitterionic lipid can complex with DNA when the interaction is mediated by a divalent cation. [9][10][11][12][13][14] Complexes formed by neutral lipids in the absence of the divalent cation have also been described. 15,16 The divalent cation-lipid complexes are entirely noncytotoxic and thereby have a remarkable status among the candidate systems in that the binding is highly tunable.…”

mentioning

confidence: 99%

Observation of a Rectangular Columnar Phase in a DNA−Calcium−Zwitterionic Lipid Complex

McManus

Rädler²,

Dawson³

2004

J. Am. Chem. Soc.

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Complexes composed of DNA and cationic lipids have been studied in detail in recent years, mainly due to their potential to deliver DNA to cells in gene therapy. 1 It has been recognized that different structures can be prepared, based on these compositions and lipid chemistry. In this communication, we give the first example of a higher-order arrangement of DNA within an entirely noncytotoxic complex.This opens the pathway to explore the consequences of such ordering on the issue of delivery and other arenas where DNA arrays may have biological significance. We believe such nontoxic ordered arrays of DNA may in the future represent novel components of structural and functional significance at the interface of the "synthetic biology" endeavor. 2 Until now most studies on these synthetic systems have been done with the focus of creating artificial devices for gene delivery. In these cases, some researchers have screened large numbers of different lipid-DNA formulations to identify successful candidates in vitro. 3 Others, however, have tried to understand the complexes on a more fundamental level, studying structure, phase behavior, and other physical characteristics, in the hope that this would ultimately enable a "design"-based approach to delivery, 4-8 but no systematic understanding has emerged.However, we consider that there emerges a general role for the engineering of ordered arrays of DNA that can protect the DNA in a biological environment but which is easily manipulated to provide compact readable and addressable devices. Such complexes typically cannot be based on direct electrostatic interaction between cationic lipids and DNA, as these are quite cytotoxic and the degree of binding is ill-controlled after initial preparation. However, complexes composed entirely of natural, zwitterionic lipid can complex with DNA when the interaction is mediated by a divalent cation. [9][10][11][12][13][14] Complexes formed by neutral lipids in the absence of the divalent cation have also been described. 15,16 The divalent cation-lipid complexes are entirely noncytotoxic and thereby have a remarkable status among the candidate systems in that the binding is highly tunable. They have been studied less, possibly due to the erroneous assumption that the interaction between divalent cations and zwitterionic lipids is weak. We have shown previously that the complex forms over a number of calcium concentrations. Structural studies of both the cationic and zwitterionic DNA lipoplexes have been reported over the past number of years. [4][5][6][13][14][15][16][17][18][19][20][21] However, here we show for the first time that, in addition to the enhanced divalent cation binding, 17 this complex is capable of arranging DNA into higher orders of local organization.The complex is prepared by mixing DNA (calf thymus, purified by a method described before 18 ), unilamellar liposomes of DPPC, prepared by bath sonication at 50°C, and calcium from concentrated stocks in the wide part of a 1 mm X-ray capillary. The complex was allowed t...

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DNA initiates polymorphic structural transitions in lecithin

Cited by 45 publications

References 25 publications

Direct visualization at the single-cell level of electrically mediated gene delivery

Direct visualization at the single-cell level of electrically mediated gene delivery

The structural diversity of DNA–neutral phospholipids–divalent metal cations aggregates: a small-angle synchrotron X-ray diffraction study

Observation of a Rectangular Columnar Phase in a DNA−Calcium−Zwitterionic Lipid Complex

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