bacmam mediated gene delivery into multipotent mesenchymal stromal cells

... the rAAV construct delivery and the transgene expression in vivo The efficiency of transgene delivery would be particularly important in diseases characterized by pan-retinal degeneration as they ... Evaluation of adeno-associated virus -mediated gene transfer into the rat retina by clinical fluorescence photography Hum Gene Ther 1999, 10:641-648 Redmond TM, Hamel CP: Genetic analysis of RPE65: from ... degeneration slow mice by gene therapy Nat Genet 2000, 25:306-310 Sarra GM, Stephens C, de Alwis M, Bainbridge JW, Smith AJ, Thrasher AJ, Ali RR: Gene replacement therapy in the retinal degeneration...

Ngày tải lên: 14/08/2014, 19:22

... mechanical DNA delivery by ultrasonic cavitation Somat Cell Mol Genet 2002, 27:115-134 Fischer AJ, Stanke JJ, Omar G, Askwith CC, Burry RW: Ultrasoundmediated gene transfer into neuronal cells J Biotechnol ... macromolecules into cells via microbubblemediated cavitation and transient disruption of the plasma membrane [37,38] As the average diameter of neuronal cells is larger than that of glial cells, it ... application, even although their efficacy for gene delivery is lower [34] Liposome -mediated gene transfer involves the fusion of synthetic lipids into the plasmid membrane, which may affect cell...

Ngày tải lên: 10/08/2014, 05:21

... retroviral genes Oncogene capture The mechanism of oncogene capture appears to be dependent on the generation of a chimeric retroviraloncogene transcript (69, 70) This suggests that the risk of oncogene ... retroviral vectors is that of insertional mutagenesis and oncogene activation Insertional mutagenesis and oncogene activation As discussed above, oncogene activation can occur either by transcription ... and insertional mutagenesis [71,72] are the two risk factors of significance in retroviral mediated gene therapy The two known instances where insertional mutagenesis/ oncogene activation has...

Ngày tải lên: 14/08/2014, 19:22

... transfer into mammalian cells and the regulation of acid-inducible genes, including aopB and katA, residing on the circular and linear chromosomes respectively, and the Tiplasmid-harbored vir genes, ... Although the occurrence of horizontal gene transfer among prokaryotes is well established, the gene transfer between bacteria and eukaryotic cells is less well elucidated The finding that ... unlinked acid-inducible genes encoding different functions in different ways 5.2 Future prospective Agrobacterium was demonstrated to be able to deliver its DNA into mammalian cells Nonetheless, the...

Ngày tải lên: 17/09/2015, 17:20

... bacterial cells containing the vir gene fusions, 100 µM acetosyringone (AS) was added into the IB media to induce the vir genes expression After grown on the IB agar plates for days, the bacterial cells ... of specific mutation genes into different genetic Agrobacterium background was conducted by electroporation in this study (Charles et al., 1994) The recipient A tumefaciens cells were grown overnight ... gene expression to low pH, but not for expression of the katA gene In addition, the katA::gfp expression level in the chvGcells grown at pH 7.0 or pH 5.5 was comparable to that in the chvG+ cells...

Ngày tải lên: 17/09/2015, 17:20

... chromosomal virulence (chv) genes Therefore the effect of these factors on Agrobacterium -mediated gene delivery into the mammalian cells was investigated 3.2.3.1 Agrobacterium vir genes are not required ... required for A tumefaciens -mediated gene delivery into mammalian cells under our assay system 3.2.3.2 T-DNA border sequences are not required for DNA transfer into mammalian cells T-DNA border sequences ... the Agrobacterium -mediated mammalian cell gene transfer 3.2.3.3.3 Combinatorial effect of mutations in chromosomal genes on DNA delivery into mammalian cells The chromosomal genes chvG, chvH and...

Ngày tải lên: 17/09/2015, 17:20

... is sensitive to gentamycin 67 2.8 General protocol for gene transfer into mammalian cells 2.8.1 Preparation of bacteria E coli, A tumefaciens and R meliloti cells were grown overnight in LB ,MG/L ... culture plates 2.8.3 Agrobacterium -mediated gene transfer into mammalian cell Mammalian cells were cocultured with A tumefaciens by adding 200 µl of bacterium cells (about 4x107 bacteria) to each ... culture After h of incubation, the cells were used for gene transfer as described 2.9 General transformation and internalization of plant cells Agrobacterium cells were grown in MG/L liquid medium...

Ngày tải lên: 17/09/2015, 17:20

... foreign gene into mammalian cells at different levels Studies on microbial pathogenesis will help us in understanding the mechanism of bacteria-based DNA delivery into the mammalian cells Being ... (T-DNA) into plant cells and integrate such foreign genetic material into the plant genome followed by the consequent gene expression to induce the tumors on plant, is the best case of horizontal gene ... its gene into mammalian cells (Kunik et al., 2001), the mechanism of such transfer needs to be understood thoroughly The efficiency of Agrobacterium mediated DNA transfer into 44 mammalian cells...

Ngày tải lên: 17/09/2015, 17:20

... for gene therapy It is impossible to obtain success in gene therapy without effective gene delivery systems that can achieve high levels of therapeutic gene expression in targeted cells Gene delivery ... which are suicide gene therapy, genetic immunotherapy, tumor suppressor gene or oncogene approaches, and anti-angiogenesis gene therapy -3- Chapter One: Introduction Suicide gene therapy is one ... Baculovirus -mediated gene silencing effects in vitro Fig 16 Quantitative analyses of baculovirus -mediated gene silencing effects in C6 cells -V- Fig 17 Quantitative analyses of baculovirus -mediated gene...

Ngày tải lên: 01/10/2015, 11:39

... surface of infected cells After budding, the virus aspartyl protease PR [8] cleaves p55, thus generating a set of smaller proteins and spacer peptides (SP) encoded by the viral pol gene during the ... development and characterization of delivery systems capable of transporting sufficient amounts of pharmacologically active agents such as CAI-peptides into the HIV-1 infected cells In cell-free systems ... passage through membranes and understand better their pharmacological behaviour For delivery of the HIV-1 CAI into human cells a bi-modular peptide was developed and constructed consisting of a transport...

Ngày tải lên: 08/08/2014, 17:20

... differentiate from mesenchymal stromal cells (MSCs) MSCs are multipotent cells that can replicate as undifferentiated cells and that have the potential to differentiate to lineages of mesenchymal tissues, ... Cells were counterstained with 4,6-diamidino-2-phenylindole (DAPI) Only irradiated cells (around 500μm in the center of plate well) were observed by fluorescence microscopy To evaluate osteogenesis, ... irradiated MSCs and controls Oteogenesis and adipogenesis of MSCs after laser irradiation Irradiating cultured MSCs with a circular beam (Fig 2A) stimulated osteogenesis exclusively within the...

Ngày tải lên: 08/08/2014, 17:20

... Adipogenesis genes are lipoprotein lipase (LPL) and proliferator-activated receptor-gamma (PPARγ), osteogenesis genes are osteocalcin (OC) and alkaline phosphatase (ALP), and chondrogenesis genes ... bone marrow mesenchymal stem cells Stem Cells 2006, 24:386-398 D'Ippolito G, Schiller PC, Ricordi C, Roos BA, Howard GA: Agerelated osteogenic potential of mesenchymal stromal stem cells from ... the species of T cells Responding peripheral blood mononuclear cells (PBMCs) (105 cells) were incubated for days with either μg/mL PHA or allogeneic stimulating PBMCs (1 × 105 cells) with or without...

Ngày tải lên: 09/08/2014, 10:23

... vector system for gene delivery Adenoviral vectors have proven efficient for gene delivery in this context [9] although high titers are required to provide sufficient levels of gene delivery to achieve ... to vascular cells including intact human saphenous vein allowing lower doses of vector to be used to achieve attractive levels of gene delivery to grafts ex vivo [13] Likewise, gene delivery to ... though the virus was only present for 2–4 weeks post grafting [11] Any improvement in gene delivery above that mediated by adenoviral serotype vectors would be very encouraging for clinical translation...

Ngày tải lên: 14/08/2014, 19:22

... MESENCHYMAL STROMAL CELLS 1.3.1 Definition of mesenchymal stromal cells 1.3.2 Heterogeneity of mesenchymal stromal cells 10 1.3.3 Homing of mesenchymal stromal cells 14 1.3.3.1 Mechanism of MSC homing ... of mesenchymal stromal cells 1.4 19 PROGRESS IN IMMUNOLOGICAL STUDY OF MESENCHYMAL STROMAL CELLS 20 1.4.1 Immunogenicity of mesenchymal stem cells 22 1.4.2 Immune modulation by mesenchymal stromal ... ARE STEM CELLS 1.2.1 Definition of stem cells 1.2.2 History of stem cells 1.2.3 Common characteristics of stem cells 1.2.4 Classification of stem cells 1.3 WHAT ARE MESENCHYMAL STROMAL CELLS 1.3.1...

Ngày tải lên: 15/09/2015, 22:09

... bone marrow cells Conclusions Our gene delivery system is specific and leads to transient gene delivery and expression It may prove useful and safe for numerous clinical applications of gene transfer ... complexes in gene delivery: from biophysics to biological applications Advanced Drug Delivery Reviews 2001, 47:277-294 Cristano RJ, Curiel DT: Strategies to gene delivery via the receptor -mediated ... plasmid into progenitor cells In the latter case, CD117+ cells were specifically transfected, and among them, Sca1+ cells were positive, suggesting a targeting of hematopietic progenitor cells...

Ngày tải lên: 14/08/2014, 19:22

... virus was generated by CreLox-driven recombination in Cre cells [22] Briefly, a confluent 10 cm2 dish of Cre cells (1.6 × 107) was split into five cm2 dishes Transfection of these cells with ... arthritis following delivery in genetically modified T cells [28] This observation suggests that targeting TGF-β1 to certain sites of inflammation through the use of arthogenic T cells also can be ... McCormack JE, Evans CH, Robbins PD: Direct retrovirus -mediated gene transfer to the synovium of the rabbit knee: implications for arthritis gene therapy Gene Ther 1997, 4:977-982 17 Mi Z, Ghivizzani...

Ngày tải lên: 09/08/2014, 01:21

... Research and Therapy 2008, 5:11 or gene- transfer vector constructs or target host genes, such as the gene for the CCR5 coreceptor [3-5] All lentivirus-based gene delivery systems contain packaging ... Rev M10 encoding gene transfer vectors We next wished to determine if the SIV RRE-based packaging system would be suitable for delivery of Rev M10 into target cells To this end, a gene transfer ... hybrid genes without the use of restriction enzymes: gene splicing by overlap extension Gene 1989, 77:61-68 Horton RM, Cai ZL, Ho SN, Pease LR: Gene splicing by overlap extension: tailor-made genes...

Ngày tải lên: 10/08/2014, 05:21

... mediate long-term transgene expression in muscle Hum Gene Ther 1997, 8:659-669 Kessler PD, Podsakoff GM, Chen X, McQuiston SA, Colosi PC, Matelis LA, Kurtzman GJ, Byrne BJ: Gene delivery to skeletal ... expression and systemic delivery of a therapeutic protein Proc Natl Acad Sci USA 1996, 93:14082-14087 Podsakoff G, Wong KK Jr, Chatterjee S: Efficient gene transfer into nondividing cells by adeno-associated ... characterization of different promoters and their respective behavior in vivo following AAV9 mediated gene delivery Our data indicate that the α-MHC promoter confers the most cardiac specific expression...

Ngày tải lên: 14/08/2014, 19:22

... SV40NLS-HMGB2186 -mediated gene delivery into COS-7 and HepG2 cells were very similar to those obtained after TATPTD-HMGB2186 -mediated transfection at comparable molar protein to DNA ratios For gene delivery into ... unidentied target molecule HMGB2186 facilitates gene delivery into COS-7 cells To investigate HMGB2186 -mediated gene transfer, the pCMS-eGFP-E2C-Luc reporter gene plasmid was used which encodes enhanced ... lM) (right panel) HMGB2186 -mediated gene transfer into COS-7 cells in the absence (C) or presence of 100 lM chloroquine (D) Cells were seeded at a density of ã 104 cells per well 24 h before...

Ngày tải lên: 16/03/2014, 22:20

... provided valuable insights into the genetic programmes utilized by M tuberculosis during dormancy adaptation [4] Transcription represents the first and the most crucial step in gene regulation in prokaryotes ... transformed into E coli XL-1 Blue The generation of the site-specific mutation was confirmed by DNA sequencing WT and K182G mutant DevR proteins were purified from E coli as described previously [11] Generation ... analysis of DevR regulon gene expression RNA was isolated from M tuberculosis cultures expressing either WT DevR protein or DevR K182G or DevR K182A mutant protein and subjected to gene expression analysis...

Ngày tải lên: 22/03/2014, 16:20