Cytoplasmic transport and nuclear import of plasmid DNA.

Published on Dec 22, 2017in Bioscience Reports2.942
· DOI :10.1042/BSR20160616
Haiqing Bai2
Estimated H-index: 2
(UR: University of Rochester),
Gillian M. Schiralli Lester2
Estimated H-index: 2
(UR: University of Rochester)
+ 1 AuthorsDavid A. Dean52
Estimated H-index: 52
(UR: University of Rochester)
Productive transfection and gene transfer require not simply the entry of DNA into cells and subsequent transcription from an appropriate promoter, but also a number of intracellular events that allow the DNA to move from the extracellular surface of the cell into and through the cytoplasm, and ultimately across the nuclear envelope and into the nucleus before any transcription can initiate. Immediately upon entry into the cytoplasm, naked DNA, either delivered through physical techniques or after disassembly of DNA–carrier complexes, associates with a large number of cellular proteins that mediate subsequent interactions with the microtubule network for movement toward the microtubule organizing center and the nuclear envelope. Plasmids then enter the nucleus either upon the mitotic disassembly of the nuclear envelope or through nuclear pore complexes in the absence of cell division, using a different set of proteins. This review will discuss our current understanding of these pathways used by naked DNA during the transfection process. While much has been elucidated on these processes, much remains to be discerned, but with the development of a number of model systems and approaches, great progress is being made.
📖 Papers frequently viewed together
89 Citations
14 Citations
#1Dollie LaJoie (HCI: Huntsman Cancer Institute)H-Index: 8
#2Katharine S. Ullman (HCI: Huntsman Cancer Institute)H-Index: 29
Each time a metazoan cell undergoes open mitosis, the nucleus is dismantled in order to partition DNA content to the daughter cells. After chromosomes separate, changes at the chromatin surface usher in reestablishment of nuclear architecture. Proteins destined for the nuclear envelope are attracted to chromatin and concomitantly recruit membrane. As nuclear envelope and protein constituents spread to coat chromatin, distinct regions emerge—some rich in rapid pore formation, others occupied by m...
32 CitationsSource
#1Svitlana Chernousova (University of Duisburg-Essen)H-Index: 12
#2Matthias Epple (University of Duisburg-Essen)H-Index: 77
The processing of DNA (for transfection) and short interfering RNA (siRNA; for gene silencing), introduced into HeLa cells by triple-shell calcium phosphate nanoparticles, was followed by live-cell imaging. For comparison, the commercial liposomal transfection agent Lipofectamine was used. The cells were incubated with these delivery systems, carrying either enhanced green fluorescent protein (eGFP)-encoding DNA or siRNA against eGFP. In the latter case, HeLa cells that stably expressed eGFP wer...
34 CitationsSource
#1Martin BeckH-Index: 47
#2Ed Hurt (Heidelberg University)H-Index: 5
Nuclear pore complexes (NPCs) are large protein assemblies that form channels in the nuclear envelope and constitute major routes for nucleocytoplasmic communication. Insights into the complex structure of NPCs provide the basis for understanding their functions and reveal how the dysfunction of their structural components, nucleoporins, contributes to human disease.
296 CitationsSource
#1Lynn GottfriedH-Index: 2
#2Xin LinH-Index: 8
Last. David A. DeanH-Index: 52
view all 4 authors...
Identification of an alveolar type I epithelial cell-specific DNA nuclear import sequence for gene delivery
7 CitationsSource
#1Mary Christie (Garvan Institute of Medical Research)H-Index: 7
#2Chiung-Wen Chang (UQ: University of Queensland)H-Index: 7
Last. Bostjan Kobe (UQ: University of Queensland)H-Index: 70
view all 11 authors...
Proteins are translated in the cytoplasm, but many need to access the nucleus to perform their functions. Understanding how these nuclear proteins are transported through the nuclear envelope and how the import processes are regulated is therefore an important aspect of understanding cell function. Structural biology has played a key role in understanding the molecular events during the transport processes and their regulation, including the recognition of nuclear targeting signals by the corres...
151 CitationsSource
#1Douglass J. Forbes (UCSD: University of California, San Diego)H-Index: 39
#2Anna Travesa (UCSD: University of California, San Diego)H-Index: 3
Last. Cyril Bernis (UCSD: University of California, San Diego)H-Index: 12
view all 4 authors...
The unexpected repurposing of nuclear transport proteins from their function in interphase to an equally vital and very different set of functions in mitosis was very surprising. The multi-talented cast when first revealed included the import receptors, importin alpha and beta, the small regulatory GTPase RanGTP, and a subset of nuclear pore proteins. In this review, we report that recent years have revealed new discoveries in each area of this expanding story in vertebrates: (a) The cast of nuc...
71 CitationsSource
#1Hidetaka Akita (Hokkaido University)H-Index: 56
#2Dai Kurihara (Hokkaido University)H-Index: 3
Last. Hideyoshi Harashima (Hokkaido University)H-Index: 79
view all 5 authors...
The nuclear transfer process is one of the critical rate-limiting processes in transgene expression. In the present study, we report on the effect of compaction and the size of the DNA molecule on nuclear transfer efficiency by microinjection. A DNA/protamine complex- or variously-sized naked DNA molecules were injected into the cytoplasm or nucleus of synchronized HeLa cells. To evaluate the nuclear transfer process, a nuclear transfer score (NT score), calculated based on transgene expression ...
15 CitationsSource
#1Jingjing Cao (ZJU: Zhejiang University)H-Index: 3
#2Cui Lin (ZJU: Zhejiang University)H-Index: 3
Last. Jiyong Zhou (ZJU: Zhejiang University)H-Index: 29
view all 8 authors...
Microtubule transport of circovirus from the periphery of the cell to the nucleus is essential for viral replication in early infection. How the microtubule is recruited to the viral cargo remains unclear. In this study, we observed that circovirus trafficking is dependent on microtubule polymerization and that incoming circovirus particles colocalize with cytoplasmic dynein and endosomes. However, circovirus binding to dynein was independent of the presence of microtubular α-tubulin and translo...
32 CitationsSource
#1Bastien CautainH-Index: 12
#2Richard Hill (University of the Algarve)H-Index: 23
Last. Wolfgang Link (University of the Algarve)H-Index: 26
view all 4 authors...
The spatial separation of DNA replication and gene transcription in the nucleus and protein translation in the cytoplasm is a uniform principle of eukaryotic cells. This compartmentalization imposes a requirement for a transport network of macromolecules to shuttle these components in and out of the nucleus. This nucleo-cytoplasmic transport of macromolecules is critical for both cell physiology and pathology. Consequently, investigating its regulation and disease-associated alterations can reve...
135 CitationsSource
#1Christelle Rosazza (University of Toulouse)H-Index: 2
#1Christelle Rosazza (University of Toulouse)H-Index: 7
Last. Marie-Pierre Rols (University of Toulouse)H-Index: 22
view all 6 authors...
Electroporation is a physical method of transferring molecules into cells and tissues. It takes advantage of the transient permeabilization of the cell membrane induced by electric field pulses, which gives hydrophilic molecules access to the cytoplasm. This method offers high transfer efficiency for small molecules that freely diffuse through electrically permeabilized membranes. Larger molecules, such as plasmid DNA, face several barriers (plasma membrane, cytoplasmic crowding, and nuclear env...
53 CitationsSource
Cited By57
#1Andy Tay (NUS: National University of Singapore)H-Index: 13
#2Nicholas A. Melosh (Stanford University)H-Index: 45
Transfection is an essential step in genetic engineering and cell therapies. While a number of non-viral micro- and nano-technologies have been developed to deliver DNA plasmids into the cell cytoplasm, one of the most challenging and least efficient steps is DNA transport to and expression in the nucleus. Here, the magnetic nano-electro-injection (MagNEI) platform is described which makes use of oscillatory mechanical stimulation after cytoplasmic delivery with high aspect-ratio nano-structures...
#1Sofía Mirón-Barroso (CSIC: Spanish National Research Council)
#2Elena B. Domènech (University of Barcelona)H-Index: 3
Last. Sonia Trigueros (CSIC: Spanish National Research Council)H-Index: 14
view all 3 authors...
Nanomaterials are currently being developed for the specific cell/tissue/organ delivery of genetic material. Nanomaterials are considered as non-viral vectors for gene therapy use. However, there are several requirements for developing a device small enough to become an efficient gene-delivery tool. Considering that the non-viral vectors tested so far show very low efficiency of gene delivery, there is a need to develop nanotechnology-based strategies to overcome current barriers in gene deliver...
#1Divya Sharma (NDSU: North Dakota State University)H-Index: 4
#2Sanjay Arora (NDSU: North Dakota State University)H-Index: 3
Last. Buddhadev Layek (NDSU: North Dakota State University)H-Index: 17
view all 4 authors...
Gene therapy encompasses the transfer of exogenous genetic materials into the patient's target cells to treat or prevent diseases. Nevertheless, the transfer of genetic material into desired cells is challenging and often requires specialized tools or delivery systems. For the past 40 years, scientists are mainly pursuing various viruses as gene delivery vectors, and the overall progress has been slow and far from the expectation. As an alternative, nonviral vectors have gained substantial atten...
#2Jorge Bolivar (UCA: University of Cádiz)H-Index: 12
view all 2 authors...
To date, vaccination has become one of the most effective strategies to control and reduce infectious diseases, preventing millions of deaths worldwide. The earliest vaccines were developed as live-attenuated or inactivated pathogens, and, although they still represent the most extended human vaccine types, they also face some issues, such as the potential to revert to a pathogenic form of live-attenuated formulations or the weaker immune response associated with inactivated vaccines. Advances i...
#1Dalinda Eusébio (UBI: University of Beira Interior)H-Index: 2
#2Ana R. Neves (UBI: University of Beira Interior)H-Index: 3
Last. Ângela Sousa (UBI: University of Beira Interior)H-Index: 10
view all 7 authors...
Abstract null null DNA vaccines have emerged as innovative approaches that have great potential to overcome the limitations of current conventional vaccines. Plasmid DNA vaccines are often safer than other vaccines because they carry only antigen genetic information, are more stable and easier to produce, and can stimulate both humoral and cellular immune responses. Although the results of ongoing clinical trials are very promising, some limitations compromise the immunogenicity of these vaccine...
#1Yi-Nan Lee (Mackay Memorial Hospital)H-Index: 3
#2Yih-Jer Wu (Mackay Medical College)H-Index: 15
Last. Hung-I Yeh (Mackay Medical College)H-Index: 28
view all 9 authors...
The therapeutic effects of ultrasonic microbubble transfection (UMT)-based vascular endothelial growth factor 165 (VEGF165) gene delivery on young and senescent endothelial progenitor cells (EPCs) were investigated. By UMT, plasmid DNA (pDNA) can be delivered into both young EPCs and senescent EPCs. In the UMT groups, higher pDNA-derived protein expression was found in senescent EPCs than in young EPCs. Consistent with this finding, a higher intracellular level of pDNA copy number was detected i...
#1Seunghyeon Shin (KAIST)H-Index: 2
#2Su Hyun Kim (KAIST)H-Index: 1
Last. Gyun Min Lee (KAIST)H-Index: 56
view all 4 authors...
A platform, based on targeted integration of transgenes using recombinase-mediated cassette exchange (RMCE) coupled with CRISPR/Cas9, is increasingly being used for the development of mammalian cell lines that produce therapeutic proteins, because of reduced clonal variation and predictable transgene expression. However, low efficiency of the RMCE process has hampered its application in multicopy or multisite integration of transgenes. To improve RMCE efficiency, nuclear transport of RMCE compon...
#1Jiahorng Liaw (TMU: Taipei Medical University)H-Index: 14
#2Wei-Hsien Hsieh (YPU: Yuanpei University)
Last. Shwu-Fen Chang (TMU: Taipei Medical University)
view all 5 authors...
Cyclo-(D-Trp-Tyr) peptide nanotubes (PNTs) were reported to be potential carriers for oral gene delivery in our previous study; however, the effect of the aspect ratio (AR) of these PNTs on gene delivery in vivo could affect penetration or interception in biological environments. The aim of this study was to assess the feasibility of cyclo-(D-Trp-Tyr) PNTs with two ARs as carriers for oral pMBP-bcl-xL-hRluc delivery to the spinal cord to treat spinal cord injury (SCI). We evaluated the biodistri...
#2Challika Kaewborisuth (Thailand National Science and Technology Development Agency)H-Index: 3
Picornaviruses are non-enveloped, single-stranded RNA viruses that cause highly contagious diseases, such as polio and hand, foot-and-mouth disease (HFMD) in human, and foot-and-mouth disease (FMD) in animals. Reverse genetics and minigenome of picornaviruses mainly depend on in vitro transcription and RNA transfection; however, this approach is inefficient due to the rapid degradation of RNA template. Although DNA-based reverse genetics systems driven by mammalian RNA polymerase I and/or II pro...
#1Gianmarco Ferri (Nest: Nest Labs)H-Index: 5
#2Giuseppe Fiume (Nest: Nest Labs)
Last. Francesco Cardarelli (Nest: Nest Labs)H-Index: 26
view all 5 authors...
Nuclear breakdown was found to be the dominant route for DNA entry into the nucleus in actively dividing cells. The possibility that alternative routes contribute to DNA entry into the nucleus, however, cannot be ruled out. Here we address the process of lipofection by monitoring the localization of fluorescently-labelled DNA plasmids at the single-cell level by confocal imaging in living interphase cells. As test formulation we choose the cationic 3β-[N-(N,N-dimethylaminoethane)-carbamoyl] chol...