H1 and HMG17 extracted from calf thymus nuclei are efficient DNA carriers in gene transfer

Zaitsev, S.; Haberland, Annekathrin; Otto, Albrecht; Vorob’ev, V. I.; Haller, Hermann; Böttger, Michael

doi:10.1038/sj.gt.3300433

Cited by 47 publications

(31 citation statements)

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“…The most active fractions identified contained histone H1 and the nonhistone protein HMG17. The nonhistone proteins HMG1 and HMG2 also may have a role as DNA carriers for gene transfer (11). Other investigators claim that histone H3 and H4 are effective in gene delivery of the HIV-1 tat gene in Jurkat cells (15); neither histone H1 nor histone H2A were effective in this study.…”

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confidence: 40%

“…Peptides corresponding to consecutive regions of charged amino acids peptides 10, 12, 13, and 16 were synthesized, and peptide 10, corresponding to residues 1-36 of histone H2A, was found to be active. Peptides that subdivided this N-terminal region (peptides [11][12][13][14] or fused it with the C terminus (peptide 17) had background levels of transfection activity. Similarly, peptide chimeras of the Nterminal portion of peptide 10 fused with sequences from H2B, H3, and H4 (peptides 27-29) lacked significant transfection activity.…”

Section: Resultsmentioning

confidence: 99%

“…It was not observed with other histones or cationic peptides (5,6). Previous studies have demonstrated that histones H1 (7)(8)(9)(10)(11)(12), H2A (5,6,13,14), and H3 and H4 (15) are effective mediators of transfection. The postulated mechanisms by which histone H1 increases gene transfection are through DNA condensation, DNase protection, and the mediation of nuclear import.…”

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confidence: 99%

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Structure and function correlation in histone H2A peptide-mediated gene transfer

Balicki

Putnam

Scaria

et al. 2002

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

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Histone H2A has been found to be efficient in DNA delivery into a number of cell lines. We have reasoned that this DNA-delivery activity is mediated by two mechanisms: (i) electrostatically driven DNA binding and condensation by histone and (ii) nuclear import of these histone H2A⅐DNA polyplexes via nuclear localization signals in the protein. We have identified a 37-aa N-terminal peptide of histone H2A that is active in in vitro gene transfer. This peptide can function as a nuclear localization signal and can bind DNA. Amino acid substitutions that replace positively charged residues and͞or DNA-binding residues of this peptide obliterate transfection activity. The introduction of a proline in the first turn of an ␣-helix of this 37-mer obliterates transfection activity, suggesting that the integrity of the ␣-helical structure of the N-terminal region of histone H2A is related to its transfection activity.transfection ͉ nuclear localization signal ͉ DNA binding G ene transfer into eukaryotic cells has the potential to treat a large number of incurable diseases. The goal of nonviral gene therapy is to mimic the successful viral mechanisms for overcoming cellular barriers that block efficient expression of the target gene while minimizing the toxicities associated with gene delivery (1). The capabilities of a synthetic nonviral vector could include specific binding to the cell surface, entry, endosomal escape, translocation to the nucleus, and in some cases stable integration into the target cell genome. The rate-limiting step of current nonviral gene delivery techniques is the transfer of encapsulated plasmids from the endosomes to the nucleus (2). The potential advantages of protein͞peptide gene transfer include ease of use, production, purity, homogeneity, ability to target nucleic acids to specific cell types, the potential for cost-effective large-scale manufacture, modular attachment of targeting ligands, and the lack of limitation on the size or type of the nucleic acid that can be delivered (1). The critical first step for efficient gene delivery is the formation of the polyplex, or complex between DNA and protein͞peptide (1-3).Eukaryotic DNA within the nucleus is packaged by wrapping around a histone octamer consisting of two molecules each of histone H2A, H2B, H3, and H4, and this packaging has dramatic effects on DNA metabolism and gene regulation (reviewed by Van Holde in ref. 4). We determined that histone octamers mediate transfection into a number of cell lines and that this activity is localized to H2A. It was not observed with other histones or cationic peptides (5, 6). Previous studies have demonstrated that histones H1 (7-12), H2A (5, 6, 13, 14), and H3 and H4 (15) are effective mediators of transfection. The postulated mechanisms by which histone H1 increases gene transfection are through DNA condensation, DNase protection, and the mediation of nuclear import. In these studies, histone H1 was considered to be the subclass of histones that mediates efficient gene transfer in the presence of chlo...

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confidence: 40%

Section: Resultsmentioning

confidence: 99%

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confidence: 99%

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Structure and function correlation in histone H2A peptide-mediated gene transfer

Balicki

Putnam

Scaria

et al. 2002

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

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“…Foreign gene expression was also improved by other DNAbinding proteins, but the greatest enhancement was obtained with the complexes that contain either NLS-H1 or calf thymus histone H1 (Fritz et al, 1996). In digitoninpermeabilized or cytosol-microinjected cells, H1 histone is imported in the cell nucleus by a facilitated transport mechanism (Breeuwer and Goldfarb, 1990;Kurz et al, 1997), and has been shown to be an efficient transfection agent, when used with a low calcium or chloroquine concentration (Chen et al, 1994;Fritz et al, 1996;Zaitsev et al, 1997;Böttger et al, 1998). Furthermore, Chen et al (1994) have developed a novel DNA delivery system to accomplish gene transfer through the asialoglycoprotein receptor-mediated endocytosis pathway.…”

Section: Histonesmentioning

confidence: 99%

“…In vivo experiments performed with liposomes/HVJ/HMG-1 complexes also revealed an efficient gene transfer in the liver (Kato et al, 1991;Tomita et al, 1992;Namiki et al, 1998). HMG1/2 or HMG17 complexes extracted from the calf thymus nuclei were also shown to be efficient DNA carriers in gene transfer (Zaitsev et al, 1997).…”

Section: Hmg-box Proteinmentioning

confidence: 99%

Improvement of exogenous DNA nuclear importation by nuclear localization signal‐bearing vectors: a promising way for non‐viral gene therapy?

Hebert

2003

Biology of the Cell

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Several vectors have been developed in order to target genes to specific cells. Virus-based vectors lead to a high transfection efficiency in vitro, but display important disadvantages such as pathological risks, which they expose to patients. Plasmid-associated chemical vectors lack these disadvantages, but allow only a very low efficiency of transgene expression. Most of the non-viral-based gene transfer techniques developed until now mainly focused their efforts to overcome the problem of DNA entry into the cell. Some recent works, however, have begun to investigate the nucleus entry problem and suggest that the trafficking of DNA from cytosol to the nucleus may be improved by using the nuclear localization signal (NLS) found in some nuclear proteins. If the vector contains one or several NLS, either as covalently or non-covalently DNA-linked peptides, a competition may take place between the rate of dissociation of the DNA-vector complexes and the rate of loading of the complexes to the NLS-mediated nucleus importation machinery. This equilibrium may be displaced towards the importation pathway by the use of NLS-bearing proteins instead of peptides. The possibility of recruiting normal endogenous cellular pathways of nuclear uptake to promote entry of exogenously applied DNA through the nuclear pore complex would, thus, seem promising. Nevertheless, attempts to improve the transport of DNA to the nucleus through the use of NLSs have achieved limited success. Although these systems show improved transgene expression, little is known about how they function in transfected cells, and the optimal formulation for gene expression is yet to be determined.

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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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H1 and HMG17 extracted from calf thymus nuclei are efficient DNA carriers in gene transfer

Cited by 47 publications

References 5 publications

Structure and function correlation in histone H2A peptide-mediated gene transfer

Structure and function correlation in histone H2A peptide-mediated gene transfer

Improvement of exogenous DNA nuclear importation by nuclear localization signal‐bearing vectors: a promising way for non‐viral gene therapy?

Recombinant mussel adhesive protein as a gene delivery material

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