A recombinant H1 histone-based system for efficient delivery of nucleic acids

Puebla, Iratxe; Esseghir, Selma; Mortlock, Alison; Brown, Aaron; Crisanti, Andrea; Low, W. P.

doi:10.1016/j.jbiotec.2003.07.006

Cited by 46 publications

(49 citation statements)

References 28 publications

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“…Physical methods, such as microinjection and electroporation, [10][11][12][13] as well as calcium co-precipitation, 14 commercially available cationic polymers and lipids, 1,15-20 and cell-penetrating peptides [21][22][23][24][25] can all be used to knock down genes. Aside from the physical methods (in which siRNAs are directly injected into the cell cytoplasm or enter via membrane pores caused by electrical bursts), all the other methods share a common characteristic complexation with a positive (cationic) charge that enables the siRNAs to interact with the negatively charged plasma membrane.…”

Section: Introductionmentioning

confidence: 99%

Special delivery: targeted therapy with small RNAs

2011

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Harnessing RNA interference using small RNA-based drugs has great potential to develop drugs designed to knock down expression of any disease-causing gene, thereby greatly expanding the universe of possible drug targets. However, delivering small RNAs into specific tissues and cells is still a hurdle. Here, we review recent progress in overcoming systemic, local and cellular barriers to RNA drug delivery, focusing on strategies for targeted uptake.

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Section: Introductionmentioning

confidence: 99%

Special delivery: targeted therapy with small RNAs

2011

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“…Two mechanisms were suggested as being responsible for this effect: (1) electrostatically driven DNA binding and condensation by histones and (2) nuclear import of histone H2A-DNA polyplexes via nuclear localization signals in the protein. Transfection of DNA in a complex with histone H1 also increases the efficiency of delivery of DNA, RNA, and small interfering RNA into immortalized and primary cells; the transfection efficiency using histone H1 was comparable to or even greater than that of liposomebased systems (Puebla et al, 2003). These authors assessed the ability of various histone H1 fragments to condense DNA and to mediate transgene delivery into primary and established neuroblastoma cells.…”

Section: Histones Can Increase Transgene Expression In Several Transfmentioning

confidence: 99%

Overexpression of SeveralArabidopsisHistone Genes IncreasesAgrobacterium-Mediated Transformation and Transgene Expression in Plants

et al. 2009

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The Arabidopsis thaliana histone H2A-1 is important for Agrobacterium tumefaciens-mediated plant transformation. Mutation of HTA1, the gene encoding histone H2A-1, results in decreased T-DNA integration into the genome of Arabidopsis roots, whereas overexpression of HTA1 increases transformation frequency. To understand the mechanism by which HTA1 enhances transformation, we investigated the effects of overexpression of numerous Arabidopsis histones on transformation and transgene expression. Transgenic Arabidopsis containing cDNAs encoding histone H2A (HTA), histone H4 (HFO), and histone H3-11 (HTR11) displayed increased transformation susceptibility, whereas histone H2B (HTB) and most histone H3 (HTR) cDNAs did not increase transformation. A parallel increase in transient gene expression was observed when histone HTA, HFO, or HTR11 overexpression constructs were cotransfected with double-or single-stranded forms of a gusA gene into tobacco (Nicotiana tabacum) protoplasts. However, these cDNAs did not increase expression of a previously integrated transgene. We identified the N-terminal 39 amino acids of H2A-1 as sufficient to increase transient transgene expression in plants. After transfection, transgene DNA accumulates more rapidly in the presence of HTA1 than with a control construction. Our results suggest that certain histones enhance transgene expression, protect incoming transgene DNA during the initial stages of transformation, and subsequently increase the efficiency of Agrobacterium-mediated transformation. INTRODUCTIONAgrobacterium tumefaciens-mediated plant genetic transformation is an important experimental tool for investigation of various aspects of plant biology and for agricultural biotechnology. Development of this transformation technology represents the culmination of many decades of effort to improve tissue culture and plant genetic engineering techniques. Agrobacteriummediated transformation is based on a conjugative transfer-like process, which eventually takes the T-DNA to the host cell nucleus (Gelvin, 2000(Gelvin, , 2003a(Gelvin, , 2009Tzfira and Citovsky, 2003;Citovsky et al., 2007). After attachment of the bacterium to plant cells and induction of Agrobacterium virulence (vir) genes, a single-stranded DNA (the T-strand) is processed from the resident tumor-inducing or root-inducing plasmid and transported from the bacterium to the plant cell. In addition, a number of Virulence effector proteins, including VirD2 (attached to the T-strand), the single-strand DNA binding protein VirE2, plus VirE3, VirD5, and VirF are also transferred to the plant (Otten et al., 1984;Stahl et al., 1998;Vergunst et al., 2000Vergunst et al., , 2003Vergunst et al., , 2005Schrammeijer et al., 2003). These proteins are likely involved in protecting T-DNA from nuclease digestion, directing T-DNA to the nucleus, stripping proteins from the T-strand prior to integration, and integrating T-DNA into the plant genome. T-DNA integration occurs randomly into genomic DNA by the process of illegitimate recombination (...

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“…Recently, various histone 1B fragments were evaluated and optimized as potential gene carriers (Puebla et al 2003). The optimized histone 1B fragment showed higher gene delivery efficiency than lipofectamine.…”

Section: Introductionmentioning

confidence: 99%

Expression, purification and characterization of TAT-high mobility group box-1A peptide as a carrier of nucleic acids

et al. 2008

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High mobility group box 1 (HMGB1) is an abundant nuclear protein that binds to double-stranded DNA. HMGB1 is composed of high mobility (HMG) box A, box B, and C-terminal acidic regions. In this study, a recombinant TAT linked HMGB1 box A (rTAT-HMGB1A) peptide was expressed, purified, and characterized as a carrier of nucleic acids. The HMGB1A cDNA was amplified by PCR, and cloned into the pET21a expression vector with the TAT domain located at the N-terminus. The rTAT-HMGB1A peptide was overexpressed and purified using Nickel affinity chromatography. A recombinant HMGB1A (rHMGB1A) peptide without the TAT domain was also overexpressed and purified as a control. In gel retardation assays, both the rHMGB1A and rTAT-HMGB1A peptides formed complexes with DNA equally well. However, transfection assays showed that the rTAT-HMGB1A peptide had a higher gene transfer efficiency than rHMGB1A. Finally, rTAT-HMGB1A had no cytotoxicity to HEK 293 cells suggesting that rTAT-HMGB1A may be useful as a non-toxic gene delivery carrier.

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A recombinant H1 histone-based system for efficient delivery of nucleic acids

Cited by 46 publications

References 28 publications

Special delivery: targeted therapy with small RNAs

Special delivery: targeted therapy with small RNAs

Overexpression of SeveralArabidopsisHistone Genes IncreasesAgrobacterium-Mediated Transformation and Transgene Expression in Plants

Expression, purification and characterization of TAT-high mobility group box-1A peptide as a carrier of nucleic acids

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