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We conclude that layered double hydroxides/DNA hybrids, like cationic lipid/DNA lipoplexes, exhibit a lamellar supraorganization with DNA molecules sandwiched between hydroxide layers, but the different chemical nature of the layers ensures the lability of these hybrid supramolec...

Self-assembly and characterization of layered double hydroxide/DNA hybrids.

NANO LETTERS, no. 2 (2006): 199-204

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摘要

The purpose of this study was to control the fabrication of new labile supramolecular assemblies by formulating associations of DNA molecules with inorganic layered double hydroxides (LDHs). The results show that LDH/DNA hybrids synthesized by a coprecipitation route involving the in situ formation of LDHs around DNA molecules acting as t...更多

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简介
  • According to the prevailing paradigm, the principle of nonviral gene delivery lies mainly in the condensation of DNA in the presence of organic cationic lipid molecules.
  • By means of X-ray diffraction, the authors sought to understand the effects of the synthesis temperature, DNA length, and hydroxide layer charge density on the structural properties of LDH/DNA hybrids.
重点内容
  • According to the prevailing paradigm, the principle of nonviral gene delivery lies mainly in the condensation of DNA in the presence of organic cationic lipid molecules
  • We report the complete characterization of magnesium-gallium layered double hydroxides (LDHs)/DNA hybrids obtained by the coprecipitation method, intercalated with double-stranded DNA fragments of various lengths (100-500 bp and 60008000 bp) and with plasmid DNA
  • We conclude that LDH/DNA hybrids, like cationic lipid/DNA lipoplexes, exhibit a lamellar supraorganization with DNA molecules sandwiched between hydroxide layers, but the different chemical nature of the layers ensures the lability of these hybrid supramolecular assemblies
  • The intercalation of DNAS into the LDH was clearly evidenced by the net increase of the interlayer distance, from ∼0.77 nm for all nitrate parent LDHs to ∼2.11 nm, ∼1.80 nm, and ∼1.96 nm for DNA molecules complexed with Mg2Al (Figure 1A), Mg2Fe (Figure 1B), and Mg2Ga (Figure 1C), respectively
  • In view of the important changes in the XRD patterns of Mg2Ga/DNAS hybrids with increasing reaction temperature and of MgRGa/DNA molecules (DNAL) hybrids with varying Mg2+/Ga3+ molar ratios, one might be tempted to say that we probably meet these extreme conditions here, at least for synthesis performed at 60 °C
  • As already mentioned for polymer-intercalated LDHs7, DNA molecules are likely to interact with charged complexes that form during cation hydrolysis, determining the Mg2+/Ga3+ molar ratio in LDH/DNA hybrids
结果
  • Of the three LDH/DNA hybrids synthesized, Mg2Ga displayed the highest signal-to-noise ratio and the least deformation of the peak profile.
  • The authors investigated whether optimal conditions designed for DNAS could be applied to long DNA molecules (DNAL), again using Mg2Ga for the hydroxide layer and 50 °C as the synthesis temperature.
  • X-ray diffraction analysis of Mg2Ga/DNAL hybrids indicated that the same d spacing (1.96 nm) was obtained irrespective of the DNA length (Figure 5).
  • The coprecipitation or self-assembly method[7,8] involves the in situ formation of LDH layers around intercalated DNA and allowed the intercalation of both short (100-500 bp) and long (6000-8000 bp) linear fragments, and plasmid DNA, into LDHs. The optimum synthesis conditions were found to be a reaction temperature of 50 °C, a magnesiumgallium LDH hydroxide layer with a Mg/Ga molar ratio of 2/1, and an amount of DNA in solution corresponding to twice the amount of Ga3+, that is, twice the theoretical anionic exchange capacity.
  • Rather well-defined materials were obtained with Mg2Ga with both short and long DNA fragments, as evidenced by the presence of several (00l) harmonics and the (110) reflection, denoting relatively good stacking of the layers and intralamellar organization.
  • In view of the important changes in the XRD patterns of Mg2Ga/DNAS hybrids with increasing reaction temperature and of MgRGa/DNAL hybrids with varying Mg2+/Ga3+ molar ratios, one might be tempted to say that the authors probably meet these extreme conditions here, at least for synthesis performed at 60 °C.
结论
  • As already mentioned for polymer-intercalated LDHs7, DNA molecules are likely to interact with charged complexes that form during cation hydrolysis, determining the Mg2+/Ga3+ molar ratio in LDH/DNA hybrids.
  • The present study describes a new selfassembly chemical approach to synthesize degradable inorganic materials at acid pH, which was made previously by anion exchange between preformed Mg2Al/NO3 and DNA.[5,12] The authors have established that by proper chemical synthesis the authors are able to entrap DNA molecules into a supramolecular assembly, which was taken up by carcinoma HeLa cells.
总结
  • According to the prevailing paradigm, the principle of nonviral gene delivery lies mainly in the condensation of DNA in the presence of organic cationic lipid molecules.
  • By means of X-ray diffraction, the authors sought to understand the effects of the synthesis temperature, DNA length, and hydroxide layer charge density on the structural properties of LDH/DNA hybrids.
  • Of the three LDH/DNA hybrids synthesized, Mg2Ga displayed the highest signal-to-noise ratio and the least deformation of the peak profile.
  • The authors investigated whether optimal conditions designed for DNAS could be applied to long DNA molecules (DNAL), again using Mg2Ga for the hydroxide layer and 50 °C as the synthesis temperature.
  • X-ray diffraction analysis of Mg2Ga/DNAL hybrids indicated that the same d spacing (1.96 nm) was obtained irrespective of the DNA length (Figure 5).
  • The coprecipitation or self-assembly method[7,8] involves the in situ formation of LDH layers around intercalated DNA and allowed the intercalation of both short (100-500 bp) and long (6000-8000 bp) linear fragments, and plasmid DNA, into LDHs. The optimum synthesis conditions were found to be a reaction temperature of 50 °C, a magnesiumgallium LDH hydroxide layer with a Mg/Ga molar ratio of 2/1, and an amount of DNA in solution corresponding to twice the amount of Ga3+, that is, twice the theoretical anionic exchange capacity.
  • Rather well-defined materials were obtained with Mg2Ga with both short and long DNA fragments, as evidenced by the presence of several (00l) harmonics and the (110) reflection, denoting relatively good stacking of the layers and intralamellar organization.
  • In view of the important changes in the XRD patterns of Mg2Ga/DNAS hybrids with increasing reaction temperature and of MgRGa/DNAL hybrids with varying Mg2+/Ga3+ molar ratios, one might be tempted to say that the authors probably meet these extreme conditions here, at least for synthesis performed at 60 °C.
  • As already mentioned for polymer-intercalated LDHs7, DNA molecules are likely to interact with charged complexes that form during cation hydrolysis, determining the Mg2+/Ga3+ molar ratio in LDH/DNA hybrids.
  • The present study describes a new selfassembly chemical approach to synthesize degradable inorganic materials at acid pH, which was made previously by anion exchange between preformed Mg2Al/NO3 and DNA.[5,12] The authors have established that by proper chemical synthesis the authors are able to entrap DNA molecules into a supramolecular assembly, which was taken up by carcinoma HeLa cells.
基金
  • This work was supported by the APEX program financed by INSERM, by an ACI program from the Ministere deleguea` la recherche et aux nouvelles technologies, and by special grants from the Association Francaise contre les Myopathies and Vaincre la Mucoviscidose
研究对象与分析
base pairs: 250
A) Powder X-ray diffraction patterns in the 2θ range 2-70° of Mg2GaNO3 (trace a) and MgRGa-DNAL hybrids prepared at 50 °C with various Mg/Ga ratios: R ) 2 (trace b), R ) 3 (trace c), R ) 4 (trace d), and R ) 5 (trace e). The vertical dashed line denotes Mg(OH)2. (B) The figure represents the corresponding in-plane reflections (110) in the 2θ range 55-65°. LDH/DNA hybrids were taken up by HeLa cells. Cells were exposed to Mg2Ga/DNA hybrids containing short DNA fragments (A, D), long DNA fragments (B, E), and plasmid DNA (C, D). DNA molecules were labeled with YOYO (1 molecule every 250 base pairs) before self-assembly with LDH. Cells were incubated with LDH/DNA hybrids for 2 h at 37 °C in 300 μL of OPTI-MEM. The medium was subsequently removed. Then, cells were washed three times with PBS and fixed with 4% paraformaldehyde for 20 min at room temperature. Finally, cells were washed with 1 mL of PBS and mounted with Vectashield with DAPI for observation. A-C correspond to the representative fluorescent micrographs. The YOYO-labeled DNA is shown in green, and nuclei are shown in blue because of the staining with DAPI. D-F correspond to the transmission images of the same cells. Magnification ×63.

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