DNA Rods Research Articles

A Novel AgI -DNA Rod Comprising a One-Dimensional Array of 11 Silver Ions within a Double Helical Structure.

DNA/RNA duplexes containing metal-ion-mediated base pairs (metallo-base pairs) have potential applications in developing nucleic acid-based nanodevices and genetic code expansion. Many metallo-base pairs are formed within duplexes stabilized by Watson-Crick base pairs. Recently, the crystal structure of an AgI -DNA nanowire with an uninterrupted one-dimensional silver array was determined. Here, we present a new DNA helical wire, the "AgI -DNA rod", containing an uninterrupted array of 11 AgI ions. The AgI -DNA rod consisted of only C-AgI -C, G-AgI -G, G-AgI -5-bromouracil (Br U), and Br U-AgI -Br U metallo base pairs, with no Watson-Crick pairs. The AgI -DNA rods were connected by non-canonical G-G pairs in crystals. Notably, data from our absorbance, circular dichroism, nuclear magnetic resonance, and mass spectrometry analyses suggested that the AgI -DNA rods formed in solution, as well as within crystals.

Chiral glass of charged DNA rods, cavity loops

Recently, the collective phase behaviors of charged DNA-viruses (rods) are explored in various low ionic strengths, where much slowing down relaxations are observed in equilibrium. These phases are distinguished by crossing the critical ionic strength, where the salt-dependent effective microscopic dynamics are important with non-monotonic multi-phase boundaries. This is also interested in terms of phase transition and replica symmetry breaking (RSB), realized by the divergence of relaxation time and the exponent power of correlation lengths in dynamic disorderorder transitions. RSB occurs as the result of many-body interactions of randomized orientations. Here, in this paper, one of the chiral mesophases, the X-pattern is highlighted for its long-time metastable state consisting of cavity loops bounded within the planar nematic, as a new type of a glass, noted as a chiral glass. The chiral glass is discussed in terms of the divergences for relaxations in the characteristic time as well the inverse of an optical pitch length, which are analyzed in the long-time-lapsed orientation textures via image-time correlations. The outcome highlights interesting aspects of charge effect in the ‘twist’ elasticity energy for optimizing the long-ranged repulsive electrostatic interactions among charged DNA rods.

Environment-Resistant DNA Origami Crystals Bridged by Rigid DNA Rods with Adjustable Unit Cells

The crystallization methodology of DNA origami frames has found salient utility in large-scalely integrating multifarious functional components following organized arrangements, thus opening up the possibilities for optical, biological, and other interdisciplinary applications. However, the single strand-dominated spacing region between adjacent DNA origami units has extremely restricted the adjustment of DNA origami separations, leading the soft crystals susceptible to environmental influences. Herein, we developed a cocrystallization pathway by incorporating rigid DNA rods into a DNA origami assembly system to achieve mutually ordered bridging on a three-dimensional scale. The intervention of DNA rods significantly improved the rigidity and crushing resistance of entire cocrystals and rendered DNA origami units exhibiting different spacing distances within the obtained crystal phase when varying DNA rod structures artificially. Such a tuning strategy that uses DNA rods as allosteric factors would provide a rational method for accessing diverse crystalline states and even modulating the tailorable properties of materials on demand.

Response of shear in bulk orientations of charged DNA rods: Taylor- and gradient-banding

Shear-induced instabilities leading to various kinds of inhomogeneous flow profiles play an important role in the processing of complex fluids, ranging from polymeric materials to various types of biological systems. In previously studied systems, either Taylor banding, or gradient banding, or fracture is observed. In the present work we study a system for which all instabilities occur in orientation textures (OTs), and where Taylor banding occurs simultaneously with gradient banding. The system here consists of crowded suspensions of long and thin DNA-based rods (at a low ionic strength of 0.16 mM salt), where the applied shear rate is systematically varied, for concentrations well below and above the glass-transition concentration (12.4 mg ml−1). To simultaneously measure the velocity profile along the gradient direction, in fracture and gradient banding, the optical cell is placed in a specially designed heterodyne light scattering set up, where the scattering volume can be scanned across the cell gap. The results confirm that Taylor bands and gradient banding occur in the concentration of DNA rods and applied shear-rates (35–80 s−1). Taylor bands clearly show the flow access in vorticity-direction, while the gradient banding is rearranged as thick rolling flows of OTs, at the middle shear-rate (50 s−1). The observations can be then useful to facilitate other biological complex fluids and the glass-forming liquids.

Equilibrium phase diagram and thermal responses of charged DNA-virus rod-suspensions at low ionic strengths

The collective behavior of DNA is important for exploring new types of bacteria in the means of detection, which is greatly interested in the understanding of interactions between DNAs in living systems. How they self-organize themselves is a physical common phenomenon for broad ranges of thermodynamic systems. In this work, the equilibrium phase diagrams of charged chiral rods (fd viruses) at low ionic strengths (below a few mM) are provided to demonstrate both replicas of (or self-organized) twist orders and replica symmetry breaking near high concentration glass-states. By varying the ionic strengths, it appears that a critical ionic strength is obtained below 1–2 mM salt, where the twist and freezing of nematic domains diverge. Also, the microscopic relaxation is revealed by the ionic strength-dependent effective Debye screening length. At a fixed low ionic strength, the local orientations of twist are shown by two different length scales of optical pitch, in the chiral-nematic N* phase and the helical domains H_{D}, for low and high concentration, respectively. RSB occurs in several cases of crossing phase boundary lines in the equilibrium phase diagram of DNA-rod concentration and ionic strength, including long-time kinetic arrests in the presence of twist orders. The different pathways of PATH I, II and III are due to many-body effects of randomized orientations for charged fd rods undergoing long-range electrostatic interactions in bulk elastic medium. In addition, the thermal stability are shown for chiral pitches of the N* phase and the abnormal cooling process of a specific heat in a structural glass. Here, the concentration-driven twist-effects of charged DNA rods are explored using various experimental methods involving image-time correlation, microscopic dynamics in small angle dynamic light scattering, optical activity in second harmonic generation, and differential scanning calorimetry for the glass state.

Engineered nano-sphere array of gold-DNA core-shells and junctions as opto-plasmonic sensors for biodetection.

In this paper, we design opto-plasmonic sensors by the engineered arrangement of gold-nanospheres. We use DNA-gold nanoparticle (GNP) core–shells and DNA rods as junctions between GNPs with a fishnet ground layer for controlling and improving the absorbance and reflection in the range of 100–300 THz. Based on available data, we check the effects of healthy and cancerous cells on the reflection parameter. Here, we demonstrate how the DNA junctions and distance between the nanospheres can be considered to modify the reflection. These structures can be utilized as opto-plasmonic sensors with high sensitivity to distinguish materials in terms of refractive indices. We can use an array of these sensors for both spectroscopy and optical imaging on a real scale. The proposed structures with different topologies are analyzed and their figure of merits (FOM) and sensitivities are obtained. The structure based on the DNA rods as junctions between GNPs shows the best FOM value of 340 RIU−1 and the core–shell heptamer structure has the best sensitivity of about 1287 nm RIU−1.

Bacterial amyloid curli acts as a carrier for DNA to elicit an autoimmune response via TLR2 and TLR9.

Bacterial biofilms are associated with numerous human infections. The predominant protein expressed in enteric biofilms is the amyloid curli, which forms highly immunogenic complexes with DNA. Infection with curli-expressing bacteria or systemic exposure to purified curli-DNA complexes triggers autoimmunity via the generation of type I interferons (IFNs) and anti-double-stranded DNA antibodies. Here, we show that DNA complexed with amyloid curli powerfully stimulates Toll-like receptor 9 (TLR9) through a two-step mechanism. First, the cross beta-sheet structure of curli is bound by cell-surface Toll-like receptor 2 (TLR2), enabling internalization of the complex into endosomes. After internalization, the curli-DNA immune complex binds strongly to endosomal TLR9, inducing production of type I IFNs. Analysis of wild-type and TLR2-deficient macrophages showed that TLR2 is the major receptor that drives the internalization of curli-DNA complexes. Suppression of TLR2 internalization via endocytosis inhibitors led to a significant decrease in Ifnβ expression. Confocal microscopy analysis confirmed that the TLR2-bound curli was required for shuttling of DNA to endosomal TLR9. Structural analysis using small-angle X-ray scattering revealed that incorporation of DNA into curli fibrils resulted in the formation of ordered curli-DNA immune complexes. Curli organizes parallel, double-stranded DNA rods at an inter-DNA spacing that matches up well with the steric size of TLR9. We also found that production of anti-double-stranded DNA autoantibodies in response to curli-DNA was attenuated in TLR2- and TLR9-deficient mice and in mice deficient in both TLR2 and TLR9 compared to wild-type mice, suggesting that both innate immune receptors are critical for shaping the autoimmune adaptive immune response. We also detected significantly lower levels of interferon-stimulated gene expression in response to purified curli-DNA in TLR2 and TLR9 deficient mice compared to wild-type mice, confirming that TLR2 and TLR9 are required for the induction of type I IFNs. Finally, we showed that curli-DNA complexes, but not cellulose, were responsible elicitation of the immune responses to bacterial biofilms. This study defines the series of events that lead to the severe pro-autoimmune effects of amyloid-expressing bacteria and suggest a mechanism by which amyloid curli acts as a carrier to break immune tolerance to DNA, leading to the activation of TLR9, production of type I IFNs, and subsequent production of autoantibodies.

Modeling DNA condensation on freestanding cationic lipid membranes

Motivated by recent experimental observations of a rapid spontaneous DNA coil-globule transition on freestanding cationic lipid bilayers, we propose simple theoretical models for DNA condensation on cationic lipid membranes. First, for a single DNA rod, we examine the conditions of full wrapping of a cylindrical DNA-like semi-flexible polyelectrolyte by an oppositely charged membrane. Then, for two parallel DNA rods, we self-consistently analyze the shape and the extent of the membrane enveloping them, focusing on membrane elastic deformations and the membrane-DNA embracing angle, which enables us to compute the membrane-mediated DNA-DNA interactions. We examine the effects of the membrane composition and its charge density, which are the experimentally tunable parameters. We show that membrane-driven rod-rod attraction is more pronounced for higher charge densities and for smaller surface tensions of the membrane. Thus, we demonstrate that for a long DNA chain adhered to a cationic lipid membrane, such membrane-induced DNA-DNA attraction can trigger compaction of DNA.

Backbone-Branched DNA Building Blocks for Facile Angular Control in Nanostructures

Nanotechnology based on the highly specific pairing of nucleobases in DNA has been used to generate a wide variety of well-defined two- and three-dimensional assemblies, both static and dynamic. However, control over the junction angles to achieve them has been limited. To achieve higher order assemblies, the strands of the DNA duplex are typically made to deviate at junctions with configurations based on crossovers or non-DNA moieties. Such strand crossovers tend to be intrinsically unstructured with the overall structural rigidity determined by the architecture of the nanoassembly, rather than the junction itself. Specific approaches to define nanoassembly junction angles are based either on the cooperative twist- and strain-promoted tuning of DNA persistence length leading to bent DNA rods for fairly large nano-objects, or de novo synthesis of individual junction inserts that are typically non-DNA and based on small organic molecules or metal-coordinating ligand moieties. Here, we describe a general strategy for direct control of junction angles in DNA nanostructures that are completely tunable about the DNA helix. This approach is used to define angular vertices through readily accessible backbone-branched DNAs (bbDNAs). We demonstrate how such bbDNAs can be used as a new building block in DNA nanoconstruction to obtain well-defined nanostructures. Angular control through readily accessible bbDNA building block provides a general and versatile approach for incorporating well-defined junctions in nanoconstructs and expands the toolkit toward achieving strain free, highly size- and shape-tunable DNA based architectures.

Structural evolution of environmentally responsive cationic liposome-DNA complexes with a reducible lipid linker.

Environmentally responsive materials (i.e., materials that respond to changes in their environment with a change in their properties or structure) are attracting increasing amounts of interest. We recently designed and synthesized a series of cleavable multivalent lipids (CMVLn, with n = 2-5 being the number of positive headgroup charges at full protonation) with a disulfide bond in the linker between their cationic headgroup and hydrophobic tails. The self-assembled complexes of the CMVLs and DNA are a prototypical environmentally responsive material, undergoing extensive structural rearrangement when exposed to reducing agents. We investigated the structural evolution of CMVL-DNA complexes at varied complex composition, temperature, and incubation time using small-angle X-ray scattering (SAXS) and wide-angle X-ray scattering (WAXS). A related lipid with a stable linker, TMVL4, was used as a control. In a nonreducing environment, CMVL-DNA complexes form the lamellar (L(α)(C)) phase, with DNA rods sandwiched between lipid bilayers. However, new self-assembled phases form when the disulfide linker is cleaved by dithiothreitol or the biologically relevant reducing agent glutathione. The released DNA and cleaved CMVL headgroups form a loosely organized phase, giving rise to a characteristic broad SAXS correlation profile. CMVLs with high headgroup charge also form condensed DNA bundles. Intriguingly, the cleaved hydrophobic tails of the CMVLs reassemble into tilted chain-ordered L(β') phases upon incubation at physiological temperature (37 °C), as indicated by characteristic WAXS peaks. X-ray scattering further reveals that two of the three phases (L(βF), L(βL), and L(βI)) constituting the L(β') phase coexist in these samples. The described system may have applications in lipid-based nanotechnologies.

Structure and internal organization of overcharged cationic-lipid/peptide/DNA self-assembly complexes

The combination of cationic lipids with cationic peptides and DNA vectors can produce synergistic effects in gene delivery to eukaryotic cells. Binary complexes of cationic lipids with DNA are well-studied whereas little information is available about the structure of the ternary lipid/peptide/DNA (LPD) complexes and mechanisms defining DNA protection and delivery. Here we use synchrotron small angle X-ray scattering and dynamic light scattering zeta-potential measurements to determine structure and the net charge of supramolecular aggregates of complexes in mixtures of plasmid DNA, cationic liposomes formed from DOTAP, plus a linear cationic ε-oligolysine with the pendant α-amino acids Leu-Tyr-Arg (LYR), ε-(LYR)K10. These ternary complexes display multilamellar structures with relatively constant separation between DOTAP bilayers, accommodating a hydrated monolayer of parallel DNA rods. The DNA–DNA distance in the complexes varies as a function of the net positive to negative (lipid+peptide)/DNA charge ratio. An explanation for the observed dependence of DNA–DNA distance on charge ratio was proposed based on general polyelectrolyte properties of non-stoichiometric polycation–DNA mixtures.

Colloidal Structure and Stability of DNA/Polycations Polyplexes Investigated by Small Angle Scattering

Polyplexes of short DNA-fragments (300 b.p., 100 nm) with tailor-made amine-based polycations of different architectures (linear and hyperbranched) were investigated in buffer solution as a function of the mixing ratio with DNA. The resulting dispersed polyplexes were characterized using small-angle neutron and X-ray scattering (SANS, SAXS) as well as cryo-TEM with respect to their mesoscopic structure and their colloidal stability. The linear polyimines form rather compact structures that have a high tendency for precipitation. In contrast, the hyperbranched polycation with enzymatic-labile pentaethylenehexamine arms (PEHA) yields polyplexes colloidally stable for months. Here the polycation coating of DNA results in a homogeneous dispersion based on a fractal network with low structural organization at low polycation amount. With increasing polycation, bundles of tens of aligned DNA rods appear that are interconnected in a fractal network with a typical correlation distance on the order of 100 nm, the average length of the DNA used. With higher organization comes a decrease in stability. The 3D network built by these beams can still exhibit some stability as long as the material concentration is large enough, but the structure collapses upon dilution. SAXS shows that the complexation does not affect the local DNA structure. Interestingly, the structural findings on the DNA polyplexes apparently correlate with the transfection efficiency of corresponding siRNA complexes. In general, these finding not only show systematic trends for the colloid stability, but may allow for rational approaches to design effective transfection carriers.

Cholesteric order in systems of helical Yukawa rods

We consider the interaction potential between two chiral rod-like colloids which consist of athin cylindrical backbone decorated with a helical charge distribution on the cylindersurface. For sufficiently slender helical rods a simple scaling expression is derived whichrelates the chiral ‘twisting’ potential to the microscopic properties of the particles, such asthe internal helical pitch, charge density and electrostatic screening parameter. To predictthe behaviour of the macroscopic cholesteric pitch of the fluid bulk phase we invoke asimple second-virial theory generalized to treat anisotropic states with weakly twisteddirector fields. It is shown that, while particles with weakly coiled helices always form acholesteric phase whose helical sense is commensurate with that of the internalhelix, more strongly coiled rods lead to the formation of a cholesteric state ofopposite sense. The correlation between the helical symmetry at the microscopic andmacroscopic scale is found to be very sensitive to the pitch of the Yukawa helix. Mixinghelical particles of sufficiently disparate length and internal pitch may give rise toa demixing of the uniform cholesteric phase into two fractions with a differentmacroscopic pitch. Our findings could be relevant to the interpretation of experimentalobservations in systems of cellulose and chitin microfibres, DNA and fd virus rods.

Torque-induced deformations of charged elastic DNA rods: thin helices, loops, and precursors of DNA supercoiling

We study the deformations of charged elastic rods under applied end forces and torques. For neutral filaments, we analyze the energetics of initial helical deformations and loop formation. We supplement this elastic approach with electrostatic energies of bent filaments and find critical conditions for buckling depending on the ionic strength of the solution. We also study force-induced loop opening, for parameters relevant for DNA. Finally, some applications of this nano-mechanical DNA model to salt-dependent onset of the DNA supercoiling are discussed.

Simulation of Self-Assembly of Cationic Lipids and DNA into Structured Complexes

We present a comprehensive set of simulations that elucidates several features of experimentally observed self-assembled structures in solutions of DNA and mixtures of neutral and cationic lipids. Our simulations are based on the Noguchi-Takasu implicit-solvent coarse-grained model of lipids as rigid trimer molecules [Noguchi, H.; Takasu, M. Phys. Rev. E 2001, 64, 041913.]. This model is extended in our work so that a certain fraction, varphi(c), of the lipids carries +1e charge and DNA molecules are introduced as uniformly charged rods. The simplified coarse-grained modeling approach provides a feasible way to study the long time-scale dynamics associated with the evolution of mesoscopically large complexes from initially disordered systems. Our simulations show that, depending on the rigidity parameter kappa(s) which governs the stiffness of the membranes, both lamellar and inverted hexagonal complexes are formed at intermediate charged lipid densities. Disordered structures are formed both when large amounts of neutral lipids (small varphi(c)) are introduced which leads to the "erosion" of the spatial order, as well as for large charge densities that result in membrane rupture. A novel phase, where DNA rods and cylindrical micelles form a two-dimensional square lattice analogous to the three-dimensional cubic NaCl-type structure, is observed in the large varphi(c) regime for very soft membrane material (kappa(s) = 0).

Transitions between Distinct Compaction Regimes in Complexes of Multivalent Cationic Lipids and DNA

Cationic lipids (CLs) have found widespread use as nonviral gene carriers (vectors), including applications in clinical trials of gene therapy. However, their observed transfection efficiencies (TEs) are inferior to those of viral vectors, providing a strong incentive for a detailed understanding of CL-DNA complex behavior. In recent systematic studies employing monovalent as well as newly synthesized multivalent lipids (MVLs), the membrane cationic charge density has been identified as a key parameter governing the TE of lamellar CL-DNA complexes. In this work, we use x-ray scattering and molecular simulations to investigate the structural properties of complexes containing MVLs. At low mole fraction of neutral lipids (NLs), Φ NL, the complexes show dramatic DNA compaction, down to essentially close-packed DNA arrays with a DNA interaxial spacing d DNA = 25 Å. A gradual increase in Φ NL does not lead to a continuous increase in d DNA as observed for DNA complexes of monovalent CLs. Instead, distinct spacing regimes exist, with sharp transitions between the regimes. Three packing states have been identified: 1), close packed, 2), condensed, but not close packed, with d DNA = 27–28 Å, and 3), an expanded state, where d DNA increases gradually with Φ NL. Based on our experimental and computational results, we conclude that the DNA condensation is mediated by the multivalent cationic lipids, which assemble between the negatively charged DNA rods. Quite remarkably, the computational results show that the less tightly packed structure in regime 2 is thermodynamically more stable than the close packed structure in regime 1. Accordingly, the constant DNA spacing observed in regime 2 is attributed to lateral phase coexistence between this stable CL-DNA complex and neutral membranes. This finding may explain the reduced TE measured for such complexes: transfection involves endosomal escape and disassembly of the complex, and these processes are inhibited by the high thermodynamic stability. Our results, which demonstrate the existence of an inverse correlation between the stability and transfection activity of lamellar CL-DNA complexes are, therefore, consistent with a recently proposed model of cellular entry.

Continuum transport model of Ogston sieving in patterned nanofilter arrays for separation of rod‐like biomolecules

This article proposes a simple computational transport model of rod-like short dsDNA molecules through a microfabricated nanofilter array. Using a nanochannel consisting of alternate deep wells and shallow slits, it is demonstrated that the complex partitioning of rod-like DNA molecules of different sizes over the nanofilter array can be well described by continuum transport theory with the orientational entropy and anisotropic transport parameters properly quantified. In this model, orientational entropy of the rod-like DNA is calculated from the equilibrium distribution of rigid cylindrical rod near the solid wall. The flux caused by entropic differences is derived from the interaction between the DNA rods and the solid channel wall during rotational diffusion. In addition to its role as an entropic barrier, the confinement of the DNA in the shallow channels also induces large changes in the effective electrophoretic mobility for longer molecules in the presence of EOF. In addition to the partitioning/selectivity of DNA molecules by the nanofilter, this model can also be used to estimate the dispersion of separated peaks. It allows for fast optimization of nanofilter separation devices, without the need of stochastic modeling techniques that are usually required.

Role of Multivalent Cations in the Self-Assembly of Phospholipid−DNA Complexes

In view of efficient and nontoxic delivery of genes to cells, complexes made of phospholipids (noncationic) and DNA are assembled through the mediation of multivalent cations. The association of lipids with DNA is explained through the charge reversal of lipid headgroups by specific adsorption of cations. The ion binding is quantified by the Gouy-Chapman-Stern theory which provides a good estimate for the minimal concentration of cations required to produce complexes. Coarse-grained Monte Carlo calculations support X-ray diffraction experiments in the sense that lipids form inverted micelles around hexagonally arranged DNA rods, with cations in between to maintain the cohesion. The complexes are more cohesive in terms of total free energy as the cation valence increases. The presented methodology may help develop predictive models for biomolecular self-assembled systems.

Terminal Twist-Induced Writhe of DNA with Intrinsic Curvature

Supercoiling of a closed circular DNA rod may result from an application of terminal twist to the DNA rod by cutting the rod, rotating one of the cut faces as the other being fixed and then sealing the cut. According to White's formula, DNA supercoiling is probably accompanied by a writhe of the DNA axis. Deduced from the elastic rod model for DNA structure, an intrinsically straight closed circular DNA rod does not writhe as subject to a terminal twist, until the number of rotation exceeds a rod-dependent threshold. By contrast, a closed circular DNA rod with intrinsic curvature writhes instantly as subject to a terminal twist. This noteworthy character in fact belongs to many intrinsically curved DNA rods. By solving the dynamic equations, the linearization of the Euler-Lagrange equations governing intrinsically curved DNA rods, this paper shows that almost every clamped-end intrinsically curved DNA rod writhes instantly when subject to a terminal twist (clamped-end DNA rods include closed circular DNA rods and topological domains of open DNA rods). In terms of physical quantities, the exceptions are identified with points in R(6) whose projections onto R(5) (through ignoring the total energy density of a rod) form a subset of a quadratic hypersurface. This paper also suggests that the terminal twist induced writhe is due to the elasticity and the clamped-end boundary conditions of the DNA rods.

Time Study of DNA Condensate Morphology: Implications Regarding the Nucleation, Growth, and Equilibrium Populations of Toroids and Rods

It is well known that multivalent cations cause free DNA in solution to condense into nanometer-scale particles with toroidal and rod-like morphologies. However, it has not been shown to what degree kinetic factors (e.g., condensate nucleation) versus thermodynamic factors (e.g., DNA bending energy) determine experimentally observed relative populations of toroids and rods. It is also not clear how multimolecular DNA toroids and rods interconvert in solution. We have conducted a series of condensation studies in which DNA condensate morphology statistics were measured as a function of time and DNA structure. Here, we show that in a typical in vitro DNA condensation reaction, the relative rod population 2 min after the initiation of condensation is substantially greater than that measured after morphological equilibrium is reached (ca. 20 min). This higher population of rods at earlier time points is consistent with theoretical studies that have suggested a favorable kinetic pathway for rod nucleation. By using static DNA loops to alter the kinetics and thermodynamics of condensation, we further demonstrate that reported increases in rod populations associated with decreasing DNA length are primarily due to a change in the thermodynamics of DNA condensation, rather than a change in the kinetics of condensate nucleation or growth. The results presented also reveal that the redistribution of DNA from rods to toroids is mediated through the exchange of DNA strands with solution.