AAV Production Using Baculovirus Expression Vector System

Sandro, Quentin; Relizani, Karima; Benchaouir, Rachid

doi:10.1007/978-1-4939-9065-8_5

Cited by 12 publications

(25 citation statements)

References 12 publications

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“…Lentiviruses' strategy is to increase the transfection rate as they rely on the machinery of the cell and can target non-dividing cells [11,17]. Finally, adeno-associated viruses have been mainly utilized for genome transduction, which targets both dividing and non-dividing cells [11,18]. Despite the advantages of these systems, viral vectors still have some safety issues to overcome, such as immunogenicity and oncogenicity [4,6].…”

Section: Introductionmentioning

confidence: 99%

PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation

et al. 2020

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Over the past decade, gene therapies have attracted much attention for the development of treatments for various conditions, including cancer, neurodegenerative diseases, protein deficiencies, and autoimmune disorders. Despite the benefits of this approach, several challenges are yet to be solved to reach clinical implementation. Some of these challenges include low transfection rates, limited stability under physiological conditions, and low specificity towards the target cells. An avenue to overcome such issues is to deliver the therapies with the aid of potent cell-penetrating vectors. Non-viral vectors, such as nanostructured materials, have been successfully tested in drug and gene delivery. Here, we propose the development and in vitro evaluation of a nanostructured cell-penetrating vehicle based on core/shell, magnetite/silver nanoparticles. A subsequent conjugation of a pH-responsive polymer was used to assure that the vehicle can carry and release circular DNA. Additionally, the translocating peptide Buforin II was conjugated with the aid of a polyether amine polymer to facilitate translocation and endosome escape. The obtained nanobioconjugates (magnetite/silver-pDMAEMA-PEA-BUFII) were characterized by UV-Vis spectrophotometry, dynamic light scattering (DLS), thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FTIR), scanning electron microscope equipped with energy dispersive spectroscopy (SEM+EDS), and transmission electron microscopy (TEM). They were also encapsulated in lecithin liposomes to form magnetoliposomes. The cell viability of Vero cells in the presence of the nanobioconjugates was above 95% and declined to 80% for the magnetoliposomes. The hemolytic tendency of nanobioconjugates and magnetoliposomes was below 10%, while the platelet aggregation approached that of the negative control (i.e., 35%). Cytoplasm coverage values of about 50% for both Vero and neuroblastoma cells confirmed significant cell penetration. Pearson’s correlation coefficients for both cell lines allowed us to estimate 20–40% colocalization of the nanobioconjugates with lysotracker green, which implied high levels of endosomal escape. The developed vehicles were also capable of loading around 16% of the added DNA and releasing such cargo with 8% efficiency. The developed nanoplatform holds a significant promise to enable highly efficient gene therapies as it overcomes some of the major issues associated with their eventual translation to the pre-clinical and clinical scale.

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Section: Introductionmentioning

confidence: 99%

PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation

et al. 2020

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“…Protein folding in the periplasm can be induced by the overexpression of two types of enzymes: Protein disulfide isomerases (PDIs), which are predominantly observed in the periplasm and catalyze disulfide bond oxidation, and peptidyl-prolyl-cis-trans isomerases (PPIs), which catalyze X–pro bond isomerization [ 138 ]. At low temperatures, co-overexpression of DsbA and the heat-shock element improves the yield of correctly folded T-cell-receptor (TCR) fragments.…”

Section: Co-expression Of Chaperonesmentioning

confidence: 99%

“…To extract soluble active proteins from inclusion bodies, the insoluble inclusion bodies must be first solubilized in denaturants ( Figure 5 ) and subsequently accompanied by a cycle of refolding (herein, referred to as the “one-step denaturing and refolding” procedure for comparison) [ 148 ]. This technique has been used for more than 20 years and is highly suitable for many bodily proteins, with approximately 40% being replenished into soluble and biologically active forms [ 138 , 149 ]. In this process, the inclusion bodies are denatured with a denaturing buffer containing either 6 M guanidine hydrochloride (GdnHCl), 8 M urea, or 0.3% sarkosyl (n-lauroyl sacosinate) [ 150 ].…”

Section: Soluble and Insoluble Expressionmentioning

confidence: 99%

Strategies for Optimizing the Production of Proteins and Peptides with Multiple Disulfide Bonds

Lee

Park

2020

Antibiotics

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Bacteria can produce recombinant proteins quickly and cost effectively. However, their physiological properties limit their use for the production of proteins in their native form, especially polypeptides that are subjected to major post-translational modifications. Proteins that rely on disulfide bridges for their stability are difficult to produce in Escherichia coli. The bacterium offers the least costly, simplest, and fastest method for protein production. However, it is difficult to produce proteins with a very large size. Saccharomyces cerevisiae and Pichia pastoris are the most commonly used yeast species for protein production. At a low expense, yeasts can offer high protein yields, generate proteins with a molecular weight greater than 50 kDa, extract signal sequences, and glycosylate proteins. Both eukaryotic and prokaryotic species maintain reducing conditions in the cytoplasm. Hence, the formation of disulfide bonds is inhibited. These bonds are formed in eukaryotic cells during the export cycle, under the oxidizing conditions of the endoplasmic reticulum. Bacteria do not have an advanced subcellular space, but in the oxidizing periplasm, they exhibit both export systems and enzymatic activities directed at the formation and quality of disulfide bonds. Here, we discuss current techniques used to target eukaryotic and prokaryotic species for the generation of correctly folded proteins with disulfide bonds.

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“…Having a bioengineered gene sequence which is potentially functioning is not enough to treat a disease. This sequence needs to be delivered to the right place in the right target cells or tissue [31]. The idea to use a virus as a transporter to insert the functional gene into these cells came after successfully using this procedure in simple goals such as transferring genetic material to a target cell [31].…”

Section: Viral Gene Deliverymentioning

confidence: 99%

“…This sequence needs to be delivered to the right place in the right target cells or tissue [31]. The idea to use a virus as a transporter to insert the functional gene into these cells came after successfully using this procedure in simple goals such as transferring genetic material to a target cell [31]. The viral gene delivery can be carried out via two paths of administration: intravitreal injection, or sub-retinal injection [9].…”

Section: Viral Gene Deliverymentioning

confidence: 99%

Trial Interventions in ABCA4- Retinopathy

Breiki

2020

Preprint

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Inherited retinal diseases collectively are one of the leading causes of visual impairment worldwide. ABCA4 retinopathy is the most common inherited retinal disease. In the last few years, there are many advances in the understanding of this disease, which led to many interventional trials with promising results at the moment. This paper is going to present a brief background up to date knowledge in the context of the disease pathophysiology, the main mechanisms involved in this disease and related diseases, and different approaches of the intervention trials. I will be using a literature review to highlight the main structural, physiological, chemical, functional characteristics of ABCA4 transporter physiology. This will be followed by a brief discussion about pathophysiology. Next, analysing, extracting and comparing different interventional options and presenting the current clinical trials for each will be done. This paper expected to discusses and rationally compare between different treatment trials approaches their advantages over others and challenges. A sensible, possible application from current trials from different diseases with similar pathogenesis and possible research opportunities will be discussed.

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AAV Production Using Baculovirus Expression Vector System

Cited by 12 publications

References 12 publications

PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation

PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation

Strategies for Optimizing the Production of Proteins and Peptides with Multiple Disulfide Bonds

Trial Interventions in ABCA4- Retinopathy

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