• Title/Summary/Keyword: Non-viral vector

Search Result 36, Processing Time 0.023 seconds

Novel Trimeric Complex for Efficient Uptake of Plasmid Vector into HepG2 Cells

  • Joo, Jong-Hyuck;Park, Jong-Gu
    • Biomedical Science Letters
    • /
    • v.9 no.2
    • /
    • pp.67-74
    • /
    • 2003
  • Viral and non-viral vectors have been used in the delivery of genetic materials into animal cells and tissues, with each approach having pros and cons. Non-viral vectors have many useful merits such as easy preparation, low immunity and size tolerance of a transgene when compared to those of viral vectors. Delivery specificity may be achieved by complex formation between receptor ligands and a non-viral vector. In the present study, non-viral vector systems are investigated in an effort to find a practical delivery means for gene therapy, Receptor-ligand interaction between transferrin-receptor and transferrin was utilized for efficient gene transfer into cancer cells. A plasmid vector, pcDNA3 (LacZ) was ligated with a small duplexed oligo fragment in which a Biotin- VN$^{TM}$ phosphoramidite was placed in the middle of the oligo. The plasmid vector labeled by biotin was then conjugated with biotin-labeled transferrin via streptavidin. This trimeric conjugates were delivered to a hepatoma cell line, HepG2. The delivery efficiency of the trimeric conjugate was 2-fold higher than that of cationic liposomes used for transfection of a plasmid vector. These results demonstrate that a plasmid vector can be efficiently transferred into cells by forming a trimeric complex of plasmid vector-linker-ligand.

  • PDF

Gene Therapy Vectors: A Current Research Insight (유전자치료를 위한 벡터 개발의 연구 동향)

  • Son, Eun-Hwa;Sohn, Eun-Soo;Pyo, Suhk-Neung
    • Journal of Pharmaceutical Investigation
    • /
    • v.34 no.5
    • /
    • pp.351-362
    • /
    • 2004
  • The basic concept underlying gene therapy is that human diseases may be treated by the transfer of genetics material into specific cells of a patient in order to correct or supplement defective genes responsible for disease development. There are several systems that can be used to transfer foreign genetic material into the human body. Both viral and non-viral vectors are developed and evaluated for delivering therapeutic genes. Viral vectors are biological systems derived from naturally evolved viruses capable of transferring their genetics materials into host cells. However, the limitations associated with viral vectors, in terms of their safety, particularly immunogenecity, and their limited capacity of transgenic materials, have encouraged researchers to increasingly focus on non-viral vectors as an alternative to viral vectors. Although non-viral vectors are less efficient than viral ones, they have the advantages of safety, simplicity of preparation and high gene encapsulation capability. This article reviews the most recent studies highlighting the advantages and the limitation of gene delivery systems focused on non-viral systems compared to viral systems.

Foamy Virus Integrase in Development of Viral Vector for Gene Therapy

  • Kim, Jinsun;Lee, Ga-Eun;Shin, Cha-Gyun
    • Journal of Microbiology and Biotechnology
    • /
    • v.30 no.9
    • /
    • pp.1273-1281
    • /
    • 2020
  • Due to the broad host suitability of viral vectors and their high gene delivery capacity, many researchers are focusing on viral vector-mediated gene therapy. Among the retroviruses, foamy viruses have been considered potential gene therapy vectors because of their non-pathogenicity. To date, the prototype foamy virus is the only retrovirus that has a high-resolution structure of intasomes, nucleoprotein complexes formed by integrase, and viral DNA. The integration of viral DNA into the host chromosome is an essential step for viral vector development. This process is mediated by virally encoded integrase, which catalyzes unique chemical reactions. Additionally, recent studies on foamy virus integrase elucidated the catalytic functions of its three distinct domains and their effect on viral pathogenicity. This review focuses on recent advancements in biochemical, structural, and functional studies of foamy virus integrase for gene therapy vector research.

Recent Advances and Future Strategy in Gene Delivery System (유전자 수송계의 현재까지의 연구동향 및 앞으로의 개발전략)

  • Choi, Woo-Jeong;Kim, Chong-Kook
    • Journal of Pharmaceutical Investigation
    • /
    • v.30 no.1
    • /
    • pp.1-12
    • /
    • 2000
  • Gene therapy is a method for the treatment of diseases with introducing the gene-engineered materials into a patient with gene-deficiency disease (e.g. cystic fibrosis) or cancer to produce a therapeutic protein in a patient's cells. Successful gene therapy requires establishing both gene expression systems and delivery systems. Viral and non-viral vectors have been used for gene delivery. Viral vectors have a high transfection efficiency, but are limited in relations to issues of safety, toxicity and immunogenecity. Non-viral vectors are easy to prepare and relatively safe. However, non-viral vectors have a low transfection efficiency. Cationic liposomes are the most available among non-viral vectors. Cationic liposomes have been used to transfect cells both in vitro and in vivo experiments. Besides, several formulations containing cationic lipid are being used in clinical trials in cases of cystic fibrosis or cancer. A crucial subject to the further development of gene delivery vectors will be a long-term gene expression with following characteristics; protecting and deliverying DNA efficiently, non-toxic and non-immunogenic, and easy to produce in large scale.

  • PDF

The Action of Hepatitis B Virus Enhancer 2-Core Gene Promoter in Non-Viral and Retroviral Vectors for Hepatocyte-Specific Expression

  • Rih, Jeong-Keun;Oh, Sang-Taek;Hwang, Deog-Su;Kim, Sun-Young;Yim, Jeong-Bin
    • BMB Reports
    • /
    • v.30 no.4
    • /
    • pp.269-273
    • /
    • 1997
  • Heptocvte-specific expression induced by Hepatitis B virus (HBV) enhancer 2-core gene promoter was examined in various hepatocyte and non-hepatocyte cell lines. using non-viral and retroviral vector systems in which chloramphenicol acetyltransferase (CAT) is used as a reporter. The non-viral plasmid containing the HBV enhancer 2-core promoter exhibited 22 and 66% of CAT activities in hepatoma cell lines. HepG2 and Hep3B, respectively when compared with CAT activity expressed by CMV promoter. The CAT activities, however. were found to be marginal in other tested hepatoma cell lines as well as mouse primary hepatocytes and non-hepatocytes. The HBV enhancer 2 located upstream the CMV promoter did not affect the CMV promoter activity nor provided hepatocyte-specific expression. Transfection of retroviral plasmid DNA containing the HBV enhancer 2-core promoter as an internal promoter exhibited high and specific CAT expression in HepG2 and Hep3B cell lines but the activity value was 5 to 10 fold lower than the non-viral plasmid with identical promoter. These results suggest that the usage of HBV enhancer 2-core promoter for liver specific expression is limited to certain vectors and hepatocyte cell lines.

  • PDF

Macromolecular Cytosolic Delivery: Cell Membranes as the Primary Obstacle

  • Larson, Gretchen M.;Lee, Kyung-Dall
    • Archives of Pharmacal Research
    • /
    • v.21 no.6
    • /
    • pp.621-628
    • /
    • 1998
  • The "evolution" of a thing, a custom, an organ is thus by no means its progressus toward a goal, even less a logical progressus by the shortest route and with the least expendit ure of force, but a succession of more or less profound, mutually independent processes of subduing, plus the resistances they encounter, the attempts at transformation for the purpose of defense and reaction, and the results of successful counteractions. The form is fluid, but the "meaning" is even more so (Friedrich W. Nietzsche).

  • PDF

Safety evaluation of gene therapy - a case study of naked DNA product

  • Ahn, Byung-Ok
    • Proceedings of the Korean Society of Toxicology Conference
    • /
    • 2003.10b
    • /
    • pp.86-86
    • /
    • 2003
  • Gene therapy is a medical intervention based on modification of the genetic material of living cells. Gene transfer usually conducted using bacterial plasmid DNA and/or virus vector to express a specific protein. Gene transfer medicinal products classified as naked nucleic acid, complexed nucleic acid or non-viral vectors, viral vector, and genetically modified cells according to biological origin.(omitted)

  • PDF

Targeted Polymeric Gene Delivery for Anti-angiogenic Tumor Therapy

  • Kim, Won-Jong;Kim, Sung-Wan
    • Macromolecular Research
    • /
    • v.15 no.2
    • /
    • pp.100-108
    • /
    • 2007
  • Gene therapy has become a promising strategy for the treatment of genetically based diseases, such as cancer, which are currently considered incurable. A major obstacle in the field of cancer gene therapy is the development of a safe and efficient delivery system for therapeutic gene transfer. Non-viral vectors have attracted great interest, as they are simple to prepare, stable, easy to modify and relatively safe compared to viral vectors. In this review, an insight into the strategies developed for polyethylenimine (PEI)-based non-viral vectors has been provide, including improvement of the polyplex properties by incorporating hydrophilic spacer, poly(ethylene glycol) (PEG). Moreover, this review will summarize the strategies for the tumor targeting. Specifically, a targeted polymeric gene delivery system, PEI-g-PEG-RGD, will be introduced as an efficient gene delivery vector for tumor therapy, including its functional analysis both in vitro and in vivo.

A Sampling Strategy for Estimating Infection Rate in Vector Mosquitoes of Mosquito-borne Bovine Viral Diseases (소 모기매개 바이러스성 질병의 Vector 감염률 추정을 위한 표본추출 전략)

  • Pak, Son-Il
    • Journal of Veterinary Clinics
    • /
    • v.29 no.1
    • /
    • pp.63-67
    • /
    • 2012
  • Mosquitoes are the vectors of a number of viral diseases in cattle, such as Akabane disease, bovine ephemeral fever, Ainovirus infection, Chuzan virus infection, and Ibaraki disease. These diseases are transmitted from an infected animal to a non-infected host via the blood feeding of the vector. In Korea, the National Veterinary Research and Quarantine Services, Ministry for Food, Agriculture, Forestry and Fisheries is responsible for planning, implementation, laboratory investigations and reporting the results of the national surveillance program for mosquito-borne bovine diseases (MBD). The surveillance program, which was started in 1993, focused to determine the seroprevalence of each disease in cattle herds in space and time. From the epidemiological point of view, more important component of the surveillance program is to monitor infection rates in vectors for specific pathogens because this information is essential for a more precise understanding the dynamics of these diseases in a given environment and for determining risk of transmission. The aim of this study was to describe and compare methods for estimation of vector infection rates using maximum likelihood (MLE) and minimum infection rate in pooled samples. Factors affecting MLE such as number of pools, pooling size and diagnostic test performance are also discussed, assuming some hypothetical sampling scenarios for MBD.

Antibody-secreting macrophages generated using CpG-free plasmid eliminate tumor cells through antibody-dependent cellular phagocytosis

  • Cha, Eun Bi;Shin, Keun Koo;Seo, Jinho;Oh, Doo-Byoung
    • BMB Reports
    • /
    • v.53 no.8
    • /
    • pp.442-447
    • /
    • 2020
  • The non-viral delivery of genes into macrophages, known as hard-to-transfect cells, is a challenge. In this study, the microporation of a CpG-free and small plasmid (pCGfd-GFP) showed high transfection efficiency, sustainable transgene expression, and good cell viability in the transfections of Raw 264.7 and primary bone marrow-derived macrophages. The non-viral method using the pCGfd vector encoding anti-EGFR single-chain Fv fused with Fc (scFv-Fc) generated the macrophages secreting anti-EGFR scFv-Fc. These macrophages effectively phagocytized tumor cells expressing EGFR through the antibody-dependent mechanism, as was proved by experiments using EGFR-knockout tumor cells. Finally, peri-tumoral injections of anti-EGFR scFv-Fc-secreting macrophages were shown to inhibit tumor growth in the xenograft mouse model.