Pigs are considered as ideal donors for xenotransplantation because they have many physiological and anatomical characteristics similar to human beings. However, antibody-mediated immunity, which includes both natural and induced antibody responses, is a major challenge for the success of pig-to-primate xenotransplantation. Various genetic modification methods help to tailor pigs to be appropriate donors for xenotransplantation. In this study, we applied transcription activator-like effector nuclease (TALEN) to knock out the porcine α-1, 3-galactosyltransferase gene GGTA1, which encodes Gal epitopes that induce hyperacute immune rejection in pig-to-human xenotransplantation. Meanwhile, human leukocyte antigen-G5 gene HLA-G5, which acts as an immunosuppressive factor, was co-transfected with TALEN into porcine fetal fibroblasts. The cell colonies of GGTA1 biallelic knockout with positive transgene for HLA-G5 were chosen as nuclear donors to generate genetic modified piglets through a single round of somatic cell nuclear transfer. As a result, we successfully obtained 20 modified piglets that were positive for GGTA1 knockout (GTKO) and half of them expressed the HLA-G5 protein. Gal epitopes on the cell membrane of GTKO/HLA-G5 piglets were completely absent. Western blotting and immunofluorescence showed that HLA-G5 was expressed in the modified piglets. Functionally, the fibroblasts from the GTKO/HLA-G5 piglets showed enhanced resistance to complement-mediated lysis ability compared with those from GTKO-only or wild-type pigs. These results indicate that the GTKO/HLA-G5 pigs could be a valuable donor model to facilitate laboratory studies and clinics for xenotransplantation.
Animals ; Animals, Genetically Modified ; Gene Knockout Techniques ; HLA Antigens ; Humans ; Nuclear Transfer Techniques ; Swine ; Transplantation, Heterologous

Animal models, particularly pigs, have come to play an important role in translational biomedical research. There have been many pig models with genetically modifications via somatic cell nuclear transfer (SCNT). However, because most transgenic pigs have been produced by random integration to date, the necessity for more exact gene-mutated models using recombinase based conditional gene expression like mice has been raised. Currently, advanced genome-editing technologies enable us to generate specific gene-deleted and -inserted pig models. In the future, the development of pig models with gene editing technologies could be a valuable resource for biomedical research.
Animals ; Gene Expression ; Gene Transfer Techniques* ; Mice ; Models, Animal ; Recombinases ; Swine

Gene therapy has been considered as a promising method for treatment of many diseases, such as acquired and genetic diseases. At present, there are two major vehicles for gene delivery including viral vectors and nonviral vectors. Viral vectors appear as high gene transfection efficiency, but some deficiencies such as inflammatory responses, recombination and mutagenesis have limited their use. On account of low pathogenicity, safety and cost-effectiveness, nonviral vectors have been attracted much attention. Cationic polymers are one of the nonviral vectors which have been widely studied. This review focuses on the structure of the cationic polymers and the interaction mechanism between the vector and DNA. We try to provide a framework for the future design and synthesis of nonviral vectors with high transfection efficiency and low toxicity for gene therapy.
Cations ; chemistry ; DNA ; genetics ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Polymers ; chemistry

Animals ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Humans ; Kidney Diseases ; genetics ; therapy

OBJECTIVETo summarize the development of gene delivery vectors in peritoneal fibrosis research and discuss the feasibility and superiority of lentiviral vectors.

DATA SOURCESThe data in this article were collected from PubMed database with relevant English articles published from 1995 to 2011.

STUDY SELECTIONArticles regarding the gene therapy in peritoneal fibrosis research using non-viral vectors, adenoviral vectors, retroviral vectors, and lentiviral vectors were selected. Data were mainly extracted from 60 articles, which are listed in the reference section of this review.

RESULTSNon-viral vector-mediated gene delivery (including naked DNA for ex vivo, oligonucleotides, ultrasound- contrast agent mediated naked gene delivery, etc.) and viral vector-mediated gene delivery (including adenovirus, helper-dependant adenovirus, and retrovirus vectors) have been successfully applied both in the mechanistic investigation and the potential prevention and treatment of peritoneal fibrosis.

CONCLUSIONSPeritoneal fibrosis is a major complication of peritoneal dialysis (PD). Recently, the wide use of the gene delivery technique made it possible to access and further research peritoneal fibrosis. The use of lentiviral vector is expected to be widely used in PD research in the future due to its advantages in gene delivery.

Drug development and preclinical trials are challenging processes and more than 80% to 90% of drug candidates fail to gain approval from the United States Food and Drug Administration. Predictive and efficient tools are required to discover high quality targets and increase the probability of success in the process of new drug development. One such solution to the challenges faced in the development of new drugs and combination therapies is the use of low-cost and experimentally manageable in vivo animal models. Since the 1980's, scientists have been able to genetically modify the mouse genome by removing or replacing a specific gene, which has improved the identification and validation of target genes of interest. Now genetically engineered mouse models (GEMMs) are widely used and have proved to be a powerful tool in drug discovery processes. This review particularly covers recent fascinating technologies for drug discovery and preclinical trials, targeted transgenesis and RNAi mouse, including application and combination of inducible system. Improvements in technologies and the development of new GEMMs are expected to guide future applications of these models to drug discovery and preclinical trials.
Animals ; Drug Discovery ; Gene Transfer Techniques ; Genome ; Mice* ; Models, Animal ; United States Food and Drug Administration

The study of viruses traditionally focused on their roles as infectious agents and as tools for understanding cell biology. Recently, however, with the development of structural biology, viruses have now been receiving particular attention in nanotechnology. By chemical methods or by gene modification, viruses have been functionalized as potential building blocks for several applications, such as drug/gene delivery vehicles, advanced vaccine vehicles, and special inorganic or organic nanomaterials. Here we highlight some of the recent progresses in the medical applications of viruses.
Biomedical Research ; trends ; Gene Transfer Techniques ; Genetic Therapy ; Nanostructures ; Nanotechnology ; trends ; Vaccines ; Viruses

The efficacy of recombinant adeno-associated virus (rAAV) vector-mediated gene delivery to the gastrointestinal tract has been paid a considerable attention over the last 10 years, since our first report on the oral gene pill strategy in Nature Medicine, even though there are still several potential obstacles for this route to overcome in order to obtain efficient gene delivery. The preclinical results of oral rAAV gene medicine are summarized in this review, and special attention is paid on its pharmacokinetic and pharmacodynamic aspects with an emphasis on drug delivery, absorption, distribution and transduction. The rAAV based vectors have been shown promising results in human clinical trials with fewer safety concerns over other gene medicines. However, the underlying mechanisms and biopharmaceutical features of oral rAAV gene medicine remain to be explored extensively and intensively to develop this novel technology as a treatment for a wider range of diseases.
Administration, Oral ; Dependovirus ; genetics ; Drug Carriers ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Genetic Vectors ; Humans

In vitro gene delivery, polyethyleneimine (PEI) has been described as one of the most efficient nonviral vector. Herein the formation mechanism of PEI/DNA complexes is elucidated. The transition phase of "bead-on-string" structure in the formation of complexes was supposed to exist through spectroscopy, electrophoresis and transmission electron microscopy (TEM) technology. The construction of PEI/DNA complexes is related closely to the characteristics of PEI and DNA plasmid. As well as the dominant electrostatic effects, the nonelectrostatic interactions were thought to be partially responsible for the presence of PEI/DNA complexes even in the high ionic strength. The surface charge of complexes particles increased with the N/P ratio, but the absolute value of zeta potential was lower at the N/P ratio of 8 and 12, perhaps attributed to the use of larger DNA plasmid. As a result, the repulsion between particles was decreased and prone to aggregate to the structure like a clustered grape-string in the solution. Interestingly, contrast to the formation behavior of complexes, the PEI/DNA complexes aggregated primarily due to hydrophobic interactions while electrostatic attractions play a little role in the complexes particles aggregation in different concentrations of salt solutions. Comparable transfection efficiency in HepG2 cells was observed for the Lipofectamine 2000 and PEI/DNA complexes at the N/P ratio of 12, and showed that larger or aggregable complexes could transfect the cells in some different mechanisms.
DNA ; genetics ; Drug Carriers ; Gene Transfer Techniques ; Genetic Therapy ; Polyethyleneimine ; chemistry ; Transfection

Animals ; Dependovirus ; genetics ; immunology ; Gene Transfer Techniques ; instrumentation ; Genetic Therapy ; instrumentation ; Genetic Vectors ; genetics ; immunology ; Humans