Rita Dias

DNA conformational dynamics in the presence of catanionic mixtures

DNA conformational behavior in the presence of non‐stoichiometric mixtures of two oppositely charged surfactants, cetyltrimethylammonium bromide and sodium octyl sulfate, was directly visualized in an aqueous solution with the use of a fluorescence microscopy technique. It was found that in the presence of cationic‐rich catanionic mixtures, DNA molecules exhibit a conformational transition from elongated coil to compact globule states. Moreover, if the catanionic mixtures form positively charged vesicles, DNA is adsorbed onto the surface of the vesicles in a collapsed globular form. When anionic‐rich catanionic mixtures are present in the solution, no change in the DNA conformational behavior was detected. Cryogenic transmission electron microscopy, as well as measurements of translational diffusion coefficients of individual DNA chains, supported our optical microscopy observations.

DNA–cationic amphiphile interactions

DNA shows strong interactions with cationic cosolutes and these have both biological and technological significance. We outline our research on various mixed systems of DNA and cationic amphiphiles including the interaction of DNA with simple cationic surfactants as well as the interaction with catanionic mixtures and positively charged catanionic vesicles. An overview from phase behavior to microstructure will be presented. We will also address DNA compaction and decompaction phenomena in different systems. Finally, simulations on DNA confinement and interaction with cationic polyions are considered.

DNA-lipid systems: A physical chemistry study

It is well known that the interaction of polyelectrolytes with oppositely charged surfactants leads to an associative phase separation; however, the phase behavior of DNA and oppositely charged surfactants is more strongly associative than observed in other systems. A precipitate is formed with very low amounts of surfactant and DNA. DNA compaction is a general phenomenon in the presence of multivalent ions and positively charged surfaces; because of the high charge density there are strong attractive ion correlation effects. Techniques like phase diagram determinations, fluorescence microscopy, and ellipsometry were used to study these systems. The interaction between DNA and catanionic mixtures (i.e., mixtures of cationic and anionic surfactants) was also investigated. We observed that DNA compacts and adsorbs onto the surface of positively charged vesicles, and that the addition of an anionic surfactant can release DNA back into solution from a compact globular complex between DNA and the cationic surfactant. Finally, DNA interactions with polycations, chitosans with different chain lengths, were studied by fluorescence microscopy, in vivo transfection assays and cryogenic transmission electron microscopy. The general conclusion is that a chitosan effective in promoting compaction is also efficient in transfection.

Stepwise Disproportionation in Polyelectrolyte Complexes

Structural properties and the topology of polyelectrolyte complexes (PECs) formed in solution have been investigated under different conditions by Monte Carlo simulations using a coarse‐grained model. The extension of individual polyions has been characterized by their radius of gyration, whereas the composition of the complexes has been investigated by their net charge and their internal topological structure by a novel analysis describing how the shorter polycations link to monomers of the longer polyanion. Conditions have been found at which the polyanion and a given number of polycations form distinguishable complexes differing in (i) the polyanion conformation and (ii) the fraction of polycations being in extended and collapsed states. Thus, at equilibrium, these PECs display a stepwise variation of the degree of intrachain disproportionation within the polyanion (also referred to as intrachain segregation), concomitant with the interchain disproportionation of the polycations, which is in agreement with previous theoretical predictions. The coexistence of the different polyelectrolyte complex structures appears, generally, at mixing ratios close to but different from charge equivalence and, as a consequence, broad polyelectrolyte size distributions are commonly obtained.

Release of DNA from surfactant complexes induced by 2-hydroxypropyl-beta-cyclodextrin.

Decompaction of DNA-CTA self-assembled complexes by 2-hydroxypropyl-beta-cyclodextrin (2-HP-beta-CD) was studied and the results were compared with beta-CD. Different degrees of 2-HP substitution (0.6, 0.8 and 1.0, respectively) were used and the decompaction was successful with all degrees of substitution. Fluorescence microscopy, steady state fluorescence spectroscopy, density and sound velocity measurements, thermal melting and circular dichroism were used. Compared to previous work using alpha- and beta-CD, the fluorescence spectroscopy results showed that the 2-HP-substituted CDs more efficiently released DNA into solution. Furthermore, dissociation of macroscopically phase separated DNA-CTA complexes was achieved upon addition of 2-HP-beta-CD and the results gave strong indications on the non-equilibrium nature of the system. The globule-to-coil transition was not found to proceed through a coexistence region, which seems to be a general phenomenon in DNA decompaction using CDs.

Polyelectrolyte condensation in bulk, at surfaces, and under confinement

In this review we discuss recent results from computer simulations based on coarse-grained polyion models representing aqueous solutions of polyelectrolytes. The focus will be directed to the conformation of the polyions and, in particular, their condensation in bulk, induced by multivalent ions and oppositely charged polyelectrolytes, at responsive surfaces and under confinement.

Interactions between DNA and surfactants

The interaction between DNA and alkyltrimethylammonium bromides of different chain lengths is studied, both on a macroscopic and on a single-molecule level. The phase maps for aqueous DNA—surfactant systems as well as some interesting salt effects are presented. Some preliminary results on the structure of DNA—surfactant complexes are also given. Studies involving DNA and surfactant mixtures are also conducted.

DNA-lipid systems. An amphiphile self-assembly and polymer-surfactant perspective

The interaction between DNA and oppositely charged surfactants has been investigated by several techniques, like fluorescence microscopy, electron microscopy, phase diagram determination, and ellipsometry. The phase behaviour is more strongly associative than that in previously studied systems. A precipitate is formed for very low amounts of surfactant and DNA. DNA compaction is a general phenomenon in the presence of multivalent ions and positively charged surfaces; because of the high charge density there are strong attractive ion correlation effects. The interaction between DNA and catanionic mixtures (i.e., mixtures of cationic and anionic surfactants) was also investigated. We observed that DNA compacts and adsorbs onto the surface of positively charged vesicles and that the addition of anionic surfactant can release free DNA back into solution from a compact globular complex between DNA and cationic surfactant. Finally, we investigated DNA interactions with polycations, chitosans with different chain lengths, by fluorescence microscopy, in vivo transfections assays and cryogenic transmission electron microscopy. The general conclusion is that a chitosan effective in promoting compaction is also efficient in transfection.

Associating polymer-surfactant systems

Some recent illustrations of the phase behavior of polymer—amphiphile systems in solution are presented. Surfactant-polymer association is demonstrated for various amphiphilic synthetic and biological polymers both on a macroscopic and on a single molecular level.

The role of excluded volume and electrostatics from coarse-grain modeling of the interaction of gemini surfactants with like-charged membranes

Cationic systems composed of lipids and/or surfactants are of paramount importance in a variety of applications. Within these, gemini have attracted particular attention, mainly due to their improved aggregation properties and to the possibility of tuning offered by the presence of a spacer. In this work, a Monte Carlo simulation study with a coarse-grained model was employed to assess the interaction of cationic gemini surfactants with a like-charged model membrane. Separating the contribution of the excluded volume and that of the electrostatic effects in the organization of gemini–lipid membranes was the first goal of this work and the role of these factors was assessed varying the concentration, the spacer length and the headgroup charge of gemini surfactants. The results provide a new insight on the organization of lipid headgroups in the vicinity of gemini surfactants. It was found that the surfactant–lipid interaction is strongly affected by the surfactant spacer length, being controlled by an overall balance between excluded volume and surfactant–lipid and surfactant–surfactant electrostatic effects. It is also seen that the out-of-plane motion of the spacer has a significant effect upon membrane organization and counterion condensation. Good agreement was found with results previously obtained from atomistic simulation.