Enhanced Intracellular Mobility and Nuclear Accumulation of DNA Plasmids Associated with a Karyophilic Protein

Mesika, Adi; Kiss, Vladimir; Brumfeld, Vlad; Ghosh, Gourisankar; Reich, Ziv

doi:10.1089/hum.2005.16.200

Cited by 106 publications

(87 citation statements)

References 45 publications

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“…29,30 In addition, several groups have shown that by complexing plasmids with proteins, such as histone proteins and nuclear factor kB, or with peptide nucleic acids (PNAs) that contain nuclear localization signals, nuclear import and expression are enhanced. [31][32][33][34] As SRF and NK3 factors are coexpressed specifically in SMCs, other cell types are unable to form an import complex on the pDNA, making the SMGA promoter a tissuespecific DTS.…”

Section: Discussionmentioning

confidence: 99%

Cell-specific nuclear import of plasmid DNA in smooth muscle requires tissue-specific transcription factors and DNA sequences

Miller

Dean

2008

Gene Ther

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Two shortcomings of nonviral gene therapy are a lack of tissue-specific targeting of vectors and low levels of gene transfer. Our laboratory has begun to address these limitations by designing plasmids that enter the nucleus of specific cell types in the absence of cell division, thereby enhancing expression in a controlled manner. We have shown that a 176 bp portion of the smooth muscle g-actin (SMGA) promoter can mediate plasmid nuclear import specifically in smooth muscle cells (SMCs). Here, we demonstrate that the binding sites for serum response factor (SRF) and NKX3-1/3-2 within this DNA nuclear targeting sequence (DTS) are required for plasmid nuclear import. Knockdown of these factors with siRNA abrogates plasmid nuclear import, indicating that they are necessary cofactors. In addition, coinjection of recombinant SRF and Nkx3.2 with the vector in TC7 epithelial cells rescues import. Finally, we show that the SRF nuclear localization sequence (NLS) is required for vector nuclear import. We propose that SRF and NKX3-1/3-2 bind the SMGA DTS in the cytoplasm, thus coating the plasmid with NLSs that mediate translocation across the nuclear pore complex. This discovery could aid in the development of more efficient nonviral vectors for gene transfer to SMCs.

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Section: Discussionmentioning

confidence: 99%

Cell-specific nuclear import of plasmid DNA in smooth muscle requires tissue-specific transcription factors and DNA sequences

Miller

Dean

2008

Gene Ther

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“…pDNA is however a large hydrophilic molecule which does not easily pass the hydrophobic central channel of the NPCs. Efforts to increase the nuclear import of pDNA through NPCs by the use of NLSs have met with limited success, although this strategy initially seemed very promising [163,164]. The use of…”

Section: Reflections On Therapeutic Strategies For the Improvement Ofmentioning

confidence: 99%

Intracellular partitioning of cell organelles and extraneous nanoparticles during mitosis

Symens

Soenen

Rejman

et al. 2012

Advanced Drug Delivery Reviews

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“…[111][112][113][114] Various synthetic and natural NLS, including viral coat proteins, increase transfection with non-viral vectors, whether or not they are covalently conjugated. 22,91,[115][116][117][118][119] Nuclear translocation is affected by several variables, including DNA conformation, the specific NLS, the number of NLS peptides conjugated to each DNA fragment, and the manner in which they are conjugated. In one study, transfection with linear DNA containing a NLS peptide failed to increase expression over plasmid DNA or linear DNA without an attached NLS.…”

Section: Nuclear Translocationmentioning

confidence: 99%

Toward Development of Artificial Viruses for Gene Therapy: A Comparative Evaluation of Viral and Non‐viral Transfection

Douglas¹

2008

Biotechnology Progress

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Problems related to the safety and efficacy of gene therapies have checked the enthusiasm once surrounding this field, though it remains a promising approach for the treatment of numerous diseases. Despite the high transfection efficiencies attainable using viral vectors, manufacturing difficulties, safety concerns, and limitations related to targeting and plasmid size have prompted considerable research into the development of non‐viral vectors. Non‐viral vectors demonstrate low toxicity, low immunogenicity, and ease of manufacture. However, they have not yet achieved the transfection efficiencies displayed by viruses. The inability to explain or predict transfection efficiencies results, in part, from insufficient understanding of the intracellular processes involved in gene delivery. Increasingly, research has been undertaken to probe the processes involved in overcoming the major obstacles to vector‐mediated transfection: (1) internalization, (2) intracellular trafficking, (3) escape to the cytosol, (4) nuclear translocation, and (5) gene transcription/expression. This paper reviews and compares the pathways and techniques involved in successful viral and non‐viral transfection. In addition, this review provides evidence that non‐viral vector development has been pursued successfully thus far, producing systems capable of evading almost all major obstacles to transfection. Evaluating the abilities of non‐viral and viral vectors to overcome specific cellular barriers reveals that the greatest advantage of viral vectors may be related to viral DNA, which is transcribed considerably more efficiently than plasmid DNA. Further study in this area should enable the development of non‐viral vectors that transfect as efficiently as viral vectors.

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Enhanced Intracellular Mobility and Nuclear Accumulation of DNA Plasmids Associated with a Karyophilic Protein

Cited by 106 publications

References 45 publications

Cell-specific nuclear import of plasmid DNA in smooth muscle requires tissue-specific transcription factors and DNA sequences

Cell-specific nuclear import of plasmid DNA in smooth muscle requires tissue-specific transcription factors and DNA sequences

Intracellular partitioning of cell organelles and extraneous nanoparticles during mitosis

Toward Development of Artificial Viruses for Gene Therapy: A Comparative Evaluation of Viral and Non‐viral Transfection

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