Evaluation of Plasmid DNA for in Vivo Gene Therapy: Factors Affecting the Number of Transfected Fibers

1998 ◽  
Vol 87 (6) ◽  
pp. 763-768 ◽  
Author(s):  
Dominic J. Wells ◽  
Jake Maule ◽  
Jill McMahon ◽  
Roger Mitchell ◽  
Elsie Damien ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Plasmid Dna ◽  
Factors Affecting

scholarly journals Osteogenic differentiation as a result of BMP-2 plasmid DNA based gene therapy in vitro and in vivo

2011 ◽  
Vol 21 ◽  
pp. 230-242 ◽  
Author(s):  
F Wegman ◽  
◽  
A Bijenhof ◽  
L Schuijff ◽  
FC Öner ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Osteogenic Differentiation ◽  
Plasmid Dna

scholarly journals Construction of a bicistronic proangiogenic expression vector and its application in experimental angiogenesis in vivo.

2003 ◽  
Vol 50 (3) ◽  
pp. 875-882 ◽  
Author(s):  
Maciej Małecki ◽  
Małgorzata Przybyszewska ◽  
Przemysław Janik
Keyword(s):  
Gene Therapy ◽  
Plasmid Dna ◽  
High Efficiency ◽  
Single Gene ◽  
Encephalomyocarditis Virus ◽  
Clinical State ◽  
Entry Site ◽  
Internal Ribosome Entry

Manipulation of angiogenesis in vivo is an example of successful gene therapy strategies. Overexpression of angiogenic genes like VEGF, FGF or PDGF causes new vessel formation and improves the clinical state of patients. Gene therapy is a very promising procedure but requires large amounts of pharmaceutical-grade plasmid DNA. In this regard we have constructed a bicistronic plasmid DNA vector encoding two proangiogenic factors, VEGF165 and FGF-2. The construct (pVIF) contains the internal ribosome entry site (IRES) of the encephalomyocarditis virus (ECMV) which permits both genes to be translated from a single bicistronic mRNA. The IRES sequence allows for a high efficiency of gene expression in vivo. The pVIF vector was characterized in vitro and in vivo. In vivo angiogenesis studies showed that the bicistronic vector encoding two proangiogenic factors induces the formation of new vessels significantly more than pVEGF165 or pFGF-2 alone. In our opinion the combined proangiogenic approach with VEGF165 and FGF-2 is more powerful and efficient than single gene therapy. We also postulate that IRES sequence can serve as a useful device improving efficiency of gene therapy.

scholarly journals AAV plasmid DNA simplifies liver-directed in vivo gene therapy: comparison of expression levels after plasmid DNA-, adeno-associated virus- and adenovirus-mediated liver transfection

2010 ◽  
Vol 12 (10) ◽  
pp. 810-817 ◽  
Author(s):  
Axel Doenecke ◽  
Alexander Krömer ◽  
Marcus N. Scherer ◽  
Hans-Jürgen Schlitt ◽  
Edward K. Geissler
Keyword(s):  
Gene Therapy ◽  
Plasmid Dna ◽  
Adeno Associated Virus ◽  
Expression Levels

Ultrasound/Microbubble Enhances Foreign Gene Expression in ECV304 Cells and Murine Myocardium

2004 ◽  
Vol 36 (12) ◽  
pp. 824-831 ◽  
Author(s):  
Dongping Guo ◽  
Xiaoyu Li ◽  
Ping Sun ◽  
Zhiguang Wang ◽  
Xiuying Chen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Therapy ◽  
Plasmid Dna ◽  
Transfection Efficiency ◽  
Ultrasound Irradiation ◽  
Ultrasound Contrast Agents ◽  
Foreign Gene ◽  
Exogenous Gene

Abstract Although viral vectors are efficient systems to transfer foreign genes into cells or target tissues, safety issues remain in relation to human gene therapy. Microbubbles currently used as ultrasound contrast agents have been applied in transfection of genes. This study was designed to test the transfection efficiency and the expression of exogenous gene mediated by ultrasound irradiation enhanced air filled albumin microbubbles in ECV304 cell line in vitro and the heart of the mouse in vivo. Air filled microbubbles (2.0–4.0 μm in diameter) were created by sonicating the mixture of human albumin, glucose, mannitol and special additive that was designed for stabilization. Plasmid DNA loading the reporter genes was gently mixed with microbubbles. The mixture of plasmid DNA and microbubbles was administrated to cultured ECV304 cells and BALB/c mice (tail vein injection) under different ultrasound/microbubble conditions, and then the transfection and expression efficiency were examined. The results both in vivo and in vitro demonstrated that microbubble with ultrasound irradiation could significantly elevate the exogenous gene expression as compared with microbubble or ultrasound only. Overall, the present study showed that the ultrasound-target microbubble destruction method enhanced the exogenous gene expression in vivo and in vitro, and provided a gene therapy way not only efficient but also easy to be manipulated and carried out in clinical.

SiRNA technology, the gene therapy of the future?

2008 ◽  
Vol 149 (4) ◽  
pp. 153-159 ◽  
Author(s):  
Zsuzsanna Rácz ◽  
Péter Hamar
Keyword(s):  
Gene Therapy ◽  
Short Interfering Rna ◽  
The Future ◽  
Interfering Rna

A genetikában új korszak kezdődött 17 éve, amikor a petúniában felfedezték a koszuppressziót. Később a koszuppressziót azonosították a növényekben és alacsonyabb rendű eukariótákban megfigyelt RNS-interferenciával (RNSi). Bár a növényekben ez ősi vírusellenes gazdaszervezeti védekezőmechanizmus, emlősökben az RNSi élettani szerepe még nincs teljesen tisztázva. Az RNSi-t rövid kettős szálú interferáló RNS-ek (short interfering RNA, siRNS) irányítják. A jelen cikkben összefoglaljuk az RNSi történetét és mechanizmusát, az siRNS-ek szerkezete és hatékonysága közötti összefüggéseket, a célsejtbe való bejuttatás virális és nem virális módjait. Az siRNS-ek klinikai alkalmazásának legfontosabb akadálya az in vivo alkalmazás. Bár a hidrodinamikus kezelés állatokban hatékony, embereknél nem alkalmazható. Lehetőséget jelent viszont a szervspecifikus katéterezés. A szintetizált siRNS-ek ismert mellékhatásait szintén tárgyaljuk. Bár a génterápia ezen új területén számos problémával kell szembenézni, a sikeres in vitro és in vivo kísérletek reményt jelentenek emberi betegségek siRNS-sel történő kezelésére.

Polymeric Delivery of Proteins and Plasmid DNA for Tissue Engineering and Gene Therapy

2001 ◽  
Vol 11 (1-3) ◽  
pp. 12 ◽  
Author(s):  
Thomas P. Richardson ◽  
William L. Murphy ◽  
David J. Mooney
Keyword(s):  
Tissue Engineering ◽  
Gene Therapy ◽  
Plasmid Dna ◽  
Polymeric Delivery

Aflatoxins biosynthesis, toxicity and intervention strategies: A review

2019 ◽  
Author(s):  
Chem Int
Keyword(s):  
Developing Countries ◽  
Molecular Basis ◽  
Genetic Regulation ◽  
Intervention Strategies ◽  
Historical Background ◽  
Economic Losses ◽  
Factors Affecting ◽  
Biomarkers Of Exposure ◽  
Food Commodities

Aflatoxins (AFTs) are toxic products of fungal metabolism, associated with serious health consequences and substantial economic losses to agriculture, livestock and poultry sectors, particularly in the developing countries. This review outlines the current information on AFTs in terms of historical background, classification, relative occurrence and co-existence with other mycotoxins in various food commodities. The phenomenon of aflatoxin (AFT) biosynthesis has been elucidated with reference to molecular basis, genetic regulation and factors affecting the AFT production. Moreover, the in vivo disposition kinetics, toxicological action and toxico-pathological consequences of AFTs have also been highlighted. Currently employed strategies for the detection and detoxification of AFTs, biomarkers of exposure assessment, potential economic impact and regulatory considerations regarding the AFTs have been emphasized.

Genetic Engineering of AAV Capsid Gene for Gene Therapy Application

2020 ◽  
Vol 20 (5) ◽  
pp. 321-332
Author(s):  
Yunbo Liu ◽  
Xu Zhang ◽  
Lin Yang
Keyword(s):  
Gene Therapy ◽  
Neutralizing Antibodies ◽  
Human Subjects ◽  
Genetic Diseases ◽  
Capsid Gene ◽  
Human Populations ◽  
Stable Gene ◽  
Efficient Gene ◽  
Gene Therapy Application

Adeno-associated virus (AAV) is a promising vector for in vivo gene therapy because of its excellent safety profile and ability to mediate stable gene expression in human subjects. However, there are still numerous challenges that need to be resolved before this gene delivery vehicle is used in clinical applications, such as the inability of AAV to effectively target specific tissues, preexisting neutralizing antibodies in human populations, and a limited AAV packaging capacity. Over the past two decades, much genetic modification work has been performed with the AAV capsid gene, resulting in a large number of variants with modified characteristics, rendering AAV a versatile vector for more efficient gene therapy applications for different genetic diseases.

Exosome as a Natural Gene Delivery Vector for Cancer Treatment

2020 ◽  
Vol 20 (11) ◽  
pp. 821-830
Author(s):  
Prasad Pofali ◽  
Adrita Mondal ◽  
Vaishali Londhe
Keyword(s):  
Gene Therapy ◽  
Gene Delivery ◽  
Nucleic Acids ◽  
Cancer Treatment ◽  
Intestinal Barrier ◽  
Gene Carrier ◽  
Gene Delivery Vector ◽  
Delivery Vector

Background: Current gene therapy vectors such as viral, non-viral, and bacterial vectors, which are used for cancer treatment, but there are certain safety concerns and stability issues of these conventional vectors. Exosomes are the vesicles of size 40-100 nm secreted from multivesicular bodies into the extracellular environment by most of the cell types in-vivo and in-vitro. As a natural nanocarrier, exosomes are immunologically inert, biocompatible, and can cross biological barriers like the blood-brain barrier, intestinal barrier, and placental barrier. Objective: This review focusses on the role of exosome as a carrier to efficiently deliver a gene for cancer treatment and diagnosis. The methods for loading of nucleic acids onto the exosomes, advantages of exosomes as a smart intercellular shuttle for gene delivery and therapeutic applications as a gene delivery vector for siRNA, miRNA and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and also the limitations of exosomes as a gene carrier are all reviewed in this article. Methods: Mostly, electroporation and chemical transfection are used to prepare gene loaded exosomes. Results: Exosome-mediated delivery is highly promising and advantageous in comparison to the current delivery methods for systemic gene therapy. Targeted exosomes, loaded with therapeutic nucleic acids, can efficiently promote the reduction of tumor proliferation without any adverse effects. Conclusion: In the near future, exosomes can become an efficient gene carrier for delivery and a biomarker for the diagnosis and treatment of cancer.

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