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Lucius, Hans; Haberland, Annekathrin; Зайцев, С. В.; Dallüge, Roman; Schneider, Marc; Böttger, Michael

doi:10.1023/a:1015230010230

Cited by 16 publications

(14 citation statements)

References 20 publications

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“…As shown previously, H1–DNA complexes showed transfection efficiency over a wide range of r i values [5]. Transfection started at low r i (>0.1) and reached a maximum at r i =0.5, which remained to high H1 concentration (up to r i =50 was tested) [5]. An increase in the zeta potential/streaming potential is anticipated from calculating the molar ratios between the amino groups (arginine+lysine) and DNA phosphate over the charge‐ratio range 0.08 ( r i =0.05)–24 ( r i =50).…”

Section: Resultssupporting

confidence: 68%

“…Besides the measurement of the zeta potential, which is often not possible in cell‐cultivation medium, these data can also be obtained by measurement of the streaming potential. As shown previously, H1–DNA complexes showed transfection efficiency over a wide range of r i values [5]. Transfection started at low r i (>0.1) and reached a maximum at r i =0.5, which remained to high H1 concentration (up to r i =50 was tested) [5].…”

Section: Resultssupporting

confidence: 66%

“…The centrifugation step simply accelerated the sedimentation of the aggregated material. Independent experiments revealed that 0.2–0.3 μg of DNA are sufficient to mediate transfection [5]. This was the case when the H1–DNA complex mixture got diluted, but also when the absolute DNA and H1 concentrations for complex formation were lowered [5].…”

Section: Resultsmentioning

confidence: 99%

“…Independent experiments revealed that 0.2–0.3 μg of DNA are sufficient to mediate transfection [5]. This was the case when the H1–DNA complex mixture got diluted, but also when the absolute DNA and H1 concentrations for complex formation were lowered [5]. The first‐mentioned method, namely dilution, showed higher transfection efficiency even at very low DNA concentrations compared with the second method.…”

Section: Resultsmentioning

confidence: 99%

“…In a previous paper we reported that aggregates of H1–DNA complexes formed in 0.15 M NaCl were the most transfection‐efficient part of a heterogeneous H1–DNA complex population. Soluble but condensed complexes remaining in the supernatant after low‐speed down‐centrifugation of the aggregates were inactive in transfection, although the amount of DNA present should have been sufficient to cause transfection [5].…”

Section: Introductionmentioning

confidence: 99%

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Structural aspects of histone H1–DNA complexes and their relation to transfection efficiency

Haberland

Cartier

Heuer

et al. 2005

Biotech and App Biochem

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During transfection, polycation-DNA complexes are normally diluted by the transfection medium, which often contains salt in the physiological concentration range and serum. It is not exactly known to what extent this dilution step influences the properties of the complexes, which in turn influence the transfection efficiency. In order to gain more insight into the size-structure-transfection activity relationship, we prepared histone H1-DNA complexes in NaCl solutions at various concentrations known to determine the size and structure of the resulting complexes. We characterized the complexes by physicochemical methods. Fluorescence correlation spectroscopy enabled relative measurements of complex sizes even under physiological conditions. The different appearances of the complexes were correlated with their transfection efficiency. When transfection was performed by dilution of the complexes in cell-cultivation media, the initial structure of H1-DNA complexes preformed under distinct salt conditions had no significant influence on the transfection efficiency. The dilution of the preformed complexes with cell-cultivation medium resulted in re-formation and aggregation of the complexes. The addition of the complexes to the cells without cell-cultivation medium, however, showed a direct correlation between the size of the complexes and the transfection efficiency (correlation coefficient 0.91). Small complexes did not contribute to the transfection.

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Section: Resultssupporting

confidence: 68%

Section: Resultssupporting

confidence: 66%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Structural aspects of histone H1–DNA complexes and their relation to transfection efficiency

Haberland

Cartier

Heuer

et al. 2005

Biotech and App Biochem

Self Cite

View full text Add to dashboard Cite

show abstract

Recombinant mussel adhesive protein as a gene delivery material

Hwang¹,

Kim²,

Lim³

et al. 2008

Biotech & Bioengineering

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Efficient target gene delivery into eukaryotic cells is important for biotechnological research and gene therapy. Gene delivery based on proteins, including histones, has recently emerged as a powerful non-viral DNA transfer technique. Here, we investigated the potential use of a recombinant mussel adhesive protein, hybrid fp-151, as a gene delivery material, in view of its similar basic amino acid composition to histone proteins, and cost-effective and high-level production in Escherichia coli. After confirming DNA binding affinity, we transfected mammalian cells (human 293T and mouse NIH/3T3) with foreign genes using hybrid fp-151 as the gene delivery carrier. Hybrid fp-151 displayed comparable transfection efficiency in both mammalian cell lines, compared to the widely used transfection agent, Lipofectamine 2000. Our results indicate that this mussel adhesive protein may be used as a potential protein-based gene-transfer mediator.

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A simple and effective sample preparation method for atomic force microscopy visualization of individual DNA molecules in situ

et al. 2010

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A simple, controllable and effective sample preparation method was established for atomic force microscopy (AFM) imaging of individual DNA molecules in aqueous solution. Firstly, magnesium ion (Mg(2+)) at a concentration of 5.0-10.0 mM as a positively charged bridge was transferred onto mica to immobilize DNA molecules. Then Mg(2+)-modified mica was used to investigate DNA molecules in any buffer without magnesium ion by AFM. AFM images demonstrated that DNA molecules can be successfully observed in solution with good resolution, reproducibility, and stability. Further, this DNA sample preparation method makes AFM successful to investigate DNA molecular interaction in situ and DNA/chitosan complex in gene delivery.

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Cited by 16 publications

References 20 publications

Structural aspects of histone H1–DNA complexes and their relation to transfection efficiency

Structural aspects of histone H1–DNA complexes and their relation to transfection efficiency

Recombinant mussel adhesive protein as a gene delivery material

A simple and effective sample preparation method for atomic force microscopy visualization of individual DNA molecules in situ

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