scholarly journals Characterization of the GBoV1 Capsid and Its Antibody Interactions

Viruses ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 330
Author(s):  
Jennifer Chun Yu ◽  
Mario Mietzsch ◽  
Amriti Singh ◽  
Alberto Jimenez Ybargollin ◽  
Shweta Kailasan ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Structural Similarity ◽  
Cell Types ◽  
Cross Reactivity ◽  
Transduction Efficiency ◽  
Human Bocavirus ◽  
Fold Axis ◽  
Genome Packaging ◽  
Efficient Gene ◽  
Human Sera

Human bocavirus 1 (HBoV1) has gained attention as a gene delivery vector with its ability to infect polarized human airway epithelia and 5.5 kb genome packaging capacity. Gorilla bocavirus 1 (GBoV1) VP3 shares 86% amino acid sequence identity with HBoV1 but has better transduction efficiency in several human cell types. Here, we report the capsid structure of GBoV1 determined to 2.76 Å resolution using cryo-electron microscopy (cryo-EM) and its interaction with mouse monoclonal antibodies (mAbs) and human sera. GBoV1 shares capsid surface morphologies with other parvoviruses, with a channel at the 5-fold symmetry axis, protrusions surrounding the 3-fold axis and a depression at the 2-fold axis. A 2/5-fold wall separates the 2-fold and 5-fold axes. Compared to HBoV1, differences are localized to the 3-fold protrusions. Consistently, native dot immunoblots and cryo-EM showed cross-reactivity and binding, respectively, by a 5-fold targeted HBoV1 mAb, 15C6. Surprisingly, recognition was observed for one out of three 3-fold targeted mAbs, 12C1, indicating some structural similarity at this region. In addition, GBoV1, tested against 40 human sera, showed the similar rates of seropositivity as HBoV1. Immunogenic reactivity against parvoviral vectors is a significant barrier to efficient gene delivery. This study is a step towards optimizing bocaparvovirus vectors with antibody escape properties.

scholarly journals High-Throughput Platform for Efficient Chemical Transfection, Virus Packaging, and Transduction

Micromachines ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 387
Author(s):  
Jianxiong Zhang ◽  
Yawei Hu ◽  
Xiaoqing Wang ◽  
Peng Liu ◽  
Xiaofang Chen
Keyword(s):  
Gene Delivery ◽  
High Throughput ◽  
Large Scale ◽  
Cell Types ◽  
Long Term Storage ◽  
Efficient Gene ◽  
Genetic Modifications ◽  
Media Change ◽  
Virus Packaging ◽  
Nih 3T3

Intracellular gene delivery is normally required to study gene functions. A versatile platform able to perform both chemical transfection and viral transduction to achieve efficient gene modification in most cell types is needed. Here we demonstrated that high throughput chemical transfection, virus packaging, and transduction can be conducted efficiently on our previously developed superhydrophobic microwell array chip (SMAR-chip). A total of 169 chemical transfections were successfully performed on the chip in physically separated microwells through a few simple steps, contributing to the convenience of DNA delivery and media change on the SMAR-chip. Efficiencies comparable to the traditional transfection in multi-well plates (~65%) were achieved while the manual operations were largely reduced. Two transfection procedures, the dry method amenable for the long term storage of the transfection material and the wet method for higher efficiencies were developed. Multiple transfections in a scheduled manner were performed to further increase the transfection efficiencies or deliver multiple genes at different time points. In addition, high throughput virus packaging integrated with target cell transduction were also proved which resulted in a transgene expression efficiency of >70% in NIH 3T3 cells. In summary, the SMAR-chip based high throughput gene delivery is efficient and versatile, which can be used for large scale genetic modifications in a variety of cell types.

Freeze-thaw increases adeno-associated virus transduction of cells

2006 ◽  
Vol 291 (2) ◽  
pp. C386-C392 ◽  
Author(s):  
Sifeng Chen ◽  
Clive Wasserfall ◽  
Matthias H. Kapturczak ◽  
Mark Atkinson ◽  
Anupam Agarwal
Keyword(s):  
Endothelial Cells ◽  
Gene Delivery ◽  
Cell Types ◽  
Transduction Efficiency ◽  
Adeno Associated Virus ◽  
Cold Temperatures ◽  
Aortic Endothelial Cells ◽  
Freeze Thaw ◽  
Proximal Tubular Epithelial Cells ◽  
Renal Proximal Tubular

A combination of gene and cell-based therapies may provide significant advantages over existing treatments in terms of their effectiveness. However, long-term efficient gene delivery has been difficult to achieve in many cell types, including endothelial cells. We developed a freeze-thaw technique which significantly increases the transduction efficiency of recombinant adeno-associated virus vectors in human aortic endothelial cells (23-fold) and in human renal proximal tubular epithelial cells (128-fold) in comparison to current methods for transduction. Freeze-thaw resulted in a transient but significant increase in cell surface area by 1,174 ± 69.8 μM2per cell. Reduction of cryogenic medium volume and repeated freeze-thaw further increased transduction efficiency by 2.8- and 2.4-fold, respectively. Trypsinization, dimethylsulfoxide, and cold temperatures, which are also involved in cell preservation, had no significant impact on transduction efficiency. Increased transduction was also observed in mesenchymal stem cells (42-fold) by the freeze-thaw method. The potential mechanism of this novel technique likely involves an increase in the net permeable area of biological membranes caused by water crystallization. These findings provide a new approach for gene delivery in various cell types, particularly in those resistant to transduction by conventional methods.

scholarly journals Serotype-based evaluation of an optogenetic construct in rat cortical astrocytes

2021 ◽  
Author(s):  
Lakshmini Balachandar ◽  
Diana Borrego ◽  
Jorge Riera
Keyword(s):  
Gene Delivery ◽  
Rat Brain ◽  
Neuronal Cell ◽  
Cell Types ◽  
Transgenic Animal ◽  
Transduction Efficiency ◽  
Aav Vector ◽  
Viral Spread ◽  
Cortical Astrocytes

Optogenetics is a modern technique which has been recently expanded to non-neuronal cell types, e.g., astrocytes, and involves targeted gene delivery of light-sensitive ion channels like Channelrhodopsin-2 (ChR2). Optogenetic regulation of astrocytic activity can be used for therapeutic intervention of several neurological disorders. Astrocytic gene delivery, viz adeno-associated viral (AAV) vectors, have proven to be robust, time-, and cost-efficient contrary to the generation of transgenic animal models. When transducing astrocytes with an AAV vector, it is imperative to perform a serotype evaluation of the AAV vector due to variability in serotype transduction efficiency depending on species, target region and construct length. Rats have been a very successful animal model for studying a variety of brain disorders, from which ChR2-based intervention of astrocytes will benefit. However, the most efficient AAV capsid serotype targeting astrocytes for ChR2 expression in the in vivo rat brain cortex has not been characterized. To address this, we have evaluated AAV serotypes 1, 5, and 8 of the vector AAV-GFAP-hChR2(H134)-mCherry targeting astrocytes in the rat brain neocortex. Results show that serotype 8 exhibits promising transduction patterns, as it has demonstrated the highest tangential and radial viral spread in the rat brain. Our research will facilitate translational research for future applications of optogenetics involving the transduction of rat brain cortical astrocytes.

scholarly journals Effective transduction of osteogenic sarcoma cells by a baculovirus vector

2003 ◽  
Vol 84 (3) ◽  
pp. 697-703 ◽  
Author(s):  
Sun U. Song ◽  
Seok-Hwan Shin ◽  
Soon-Ki Kim ◽  
Gwang-Seong Choi ◽  
Woo-Chul Kim ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Cell Lines ◽  
Reporter Gene ◽  
Mammalian Cells ◽  
Osteogenic Sarcoma ◽  
Transduction Efficiency ◽  
Sarcoma Cell ◽  
Efficient Gene ◽  
High Level ◽  
Sarcoma Cells

Efficient gene delivery of a baculovirus-derived vector (BV-p53-lacZ) to a human osteogenic sarcoma cell line, Saos-2, was serendipitously found while evaluating the vector for gene delivery to human p53-null tumour cells in a previous study. Therefore, we investigated other human, rat and mouse osteogenic sarcoma and other types of tumour cell lines for transduction efficiency via baculovirus vectors containing a lacZ reporter gene under the control of either a cytomegalovirus or Rous sarcoma virus promoter. The expression of β-galactosidase protein, assessed by X-Gal staining and β-galactosidase ELISA, demonstrated an extremely high level of transduction efficiency in some osteogenic sarcoma cell lines, such as U-2OS, Saos-2 and Saos-LM2. These human osteogenic sarcoma cell lines showed levels of β-galactosidase expression 5–40 times greater than HepG2 cells, which were previously thought to be the mammalian cells most susceptible to baculovirus-mediated gene delivery. The level of acetylated histone proteins in these tumour lines did not correlate well with the high level of reporter gene expression. These results strongly suggest that some osteogenic sarcoma cells are highly susceptible to baculovirus-mediated gene delivery and that a baculovirus-derived vector is an efficient gene delivery vehicle into human osteogenic sarcoma cells.

Amino acids functionalized dendrimers with nucleus accumulation for efficient gene delivery

2021 ◽  
pp. 120641
Author(s):  
Guoxin Tan ◽  
Jiayang Li ◽  
Dandan Liu ◽  
Hao Pan ◽  
Renfang Zhu ◽  
...  
Keyword(s):  
Amino Acids ◽  
Gene Delivery ◽  
Efficient Gene

scholarly journals Carbon Dot Nanoparticles: Exploring the Potential Use for Gene Delivery in Ophthalmic Diseases

Nanomaterials ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 935
Author(s):  
Manas R. Biswal ◽  
Sofia Bhatia
Keyword(s):  
Gene Delivery ◽  
Therapeutic Option ◽  
Retinal Dystrophy ◽  
Retinal Cells ◽  
Adeno Associated Virus ◽  
Inherited Retinal Dystrophies ◽  
Retinal Dystrophies ◽  
Efficient Gene ◽  
New Type ◽  
Viral Nanoparticles

Ocular gene therapy offers significant potential for preventing retinal dystrophy in patients with inherited retinal dystrophies (IRD). Adeno-associated virus (AAV) based gene transfer is the most common and successful gene delivery approach to the eye. These days, many studies are using non-viral nanoparticles (NPs) as an alternative therapeutic option because of their unique properties and biocompatibility. Here, we discuss the potential of carbon dots (CDs), a new type of nanocarrier for gene delivery to the retinal cells. The unique physicochemical properties of CDs (such as optical, electronic, and catalytic) make them suitable for biosensing, imaging, drug, and gene delivery applications. Efficient gene delivery to the retinal cells using CDs depends on various factors, such as photoluminescence, quantum yield, biocompatibility, size, and shape. In this review, we focused on different approaches used to synthesize CDs, classify CDs, various pathways for the intake of gene-loaded carbon nanoparticles inside the cell, and multiple studies that worked on transferring nucleic acid in the eye using CDs.

Polyethylenimine-poly(amidoamine) dendrimer modified with l-arginines as an efficient gene delivery vector

2015 ◽  
Vol 23 (8) ◽  
pp. 726-733 ◽  
Author(s):  
Nan Young Ahn ◽  
Tae-Hun Kim ◽  
Su Jeong Song ◽  
Jeong-Mi Moon ◽  
Tai Hwan Ha ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Gene Delivery Vector ◽  
Delivery Vector ◽  
Efficient Gene

scholarly journals Efficient gene delivery and silencing of mouse and human pancreatic islets

2010 ◽  
Vol 10 (1) ◽  
pp. 28 ◽  
Author(s):  
Bruno Lefebvre ◽  
Brigitte Vandewalle ◽  
Justine Longue ◽  
Ericka Moerman ◽  
Bruno Lukowiak ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Pancreatic Islets ◽  
Human Pancreatic Islets ◽  
Efficient Gene

Efficient gene delivery using chitosan–polyethylenimine hybrid systems

2008 ◽  
Vol 3 (2) ◽  
pp. 025013 ◽  
Author(s):  
Hu-Lin Jiang ◽  
Tae-Hee Kim ◽  
You-Kyoung Kim ◽  
In-Young Park ◽  
Myung-Haing Cho ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Hybrid Systems ◽  
Efficient Gene
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