vector requirements for vascular gene transfer

... efficiency of lentivectors was only 87.9% for 293T cells, 77.1% for NIH 3T3 cells, and 63.9% for 293 cells for the same volume of vector (Fig 3A) When a third generation of lentiviral vector packaging ... colonies were counted as positive for gene transfer Transduction – studies of vector decay Cell-free viral vector- containing supernatant was incubated at 37°C for 30 min, hr, hr, or 24 hr prior ... cytometry analysis was performed to evaluate the expression of lentivirus vector- mediated gene transfer Cells were washed with PBS, and then fixed with 1% paraformaldehyde before the analysis Samples...

Ngày tải lên: 14/08/2014, 19:22

... promising vectors in the field of gene transfer for neurodegenerative diseases [1-3] The larger cloning capacity of lentiviral vectors, however, makes them more suitable for therapeutic purposes A variety ... processed days later for cell identification Two different kinds of vectors were used: vectors encoding β-Gal (103 vector particles per cell) and vectors encoding EGFP (104 vector particles per ... pseudotyped HIV1-based gene transfer vectors Gene Therapy 2000, 7:910-913 Marino MP, Luce MJ, Reiser J: Small- to large-scale production of lentivirus vectors In Lentivirus Gene Engineering Protocols...

Ngày tải lên: 14/08/2014, 19:22

... to neural precursors for lineage studies (Cepko et al., 2000), to tumor cells for therapeutic intervention, and for ex vivo gene transfer The use of retrovirus vectors for gene delivery to the ... particular condition 1.1.2 Overview of Gene Delivery Vectors Potential gene transfer vectors for mammalian cells are classified into physical, non-viral, and viral vectors Physical delivery modalities, ... however, gene therapy requires gene transfer vehicles capable of efficiently transducing tumor cells The research conducted for this thesis focused mainly on strategic development of gene transfer vectors...

Ngày tải lên: 12/09/2015, 09:10

... diseases, vascular diseases and other diseases The genes transferred include suicide genes and genes encoding cytokines, antigens, tumor suppressors, markers and receptors etc For updated information ... cells 1.1.2.2 The viral vectors Generally, there are two classes of vectors for gene delivery The first class is known as viral vector (Somia and Verma, 2000) This group of vectors is derived from ... original idea of retargeting vectors for selective gene transfer has been studied only for the purpose of improving the vector efficiency itself The results of early in vivo gene therapy trials have...

Ngày tải lên: 16/09/2015, 17:13

... is forthcoming Antiangiogenic Gene Therapy Antiangiogenic gene transfer inhibits formation of neovasculature required for tumor growth and may also act by collapsing immature tumor-associated vascular ... proposals for human therapy Hum Gene Ther 11: 1553 – 1567 98 Kay, M A., et al (2000) Evidence for gene transfer and expression of factor IX in haemophilia B patients treated with an AAV vector Nat Genet ... enhances gene transfer for ovarian cancer gene therapy approaches Gynecol Oncol 74: 227 – 234 Kanerva, A., et al (2002) Targeting adenovirus to the serotype receptor increases gene transfer efficiency...

Ngày tải lên: 22/03/2014, 11:20

... it to perform the following cycling (see Note 1): Initial step: 94°C for min; 30 cycles of: 94°C for 30 s (denaturation), 55°C for (annealing), 72°C for (extension); final step: 72°C for 10 Prepare ... genomic clones for the region and hence the gene of interest, which in turn can be used for FISH or the further genomic characterization of the gene Additional valuable information on the generation ... analyze the vector: for the GB4 panel at http://www-genome.wi.mit.edu/cgi-bin/contig/rhmapper.pl, for the G3 panel at http:www-shgc.stanford.edu, and for the TNG panel at http://wwwshgc.stanford.edu/RH/TNGindex.html...

Ngày tải lên: 11/04/2014, 10:39

... 17k genes DATASET PRE POST TX n=7 comment (a) Panelli et al R8.7 n=7 1578 genes 1578 genes q12,4D DATASET POST vehicle 637 genes n=7 n=7 reviews POST TX ttest p-value < 0.05 (b) reports 637 genes ... [61] Gene ratios are presented according to the central method for display [62] Gene annotations were mined using web-based tools such as DAVID [63], GeneCards [64], COPE [65] and Bioinformatic ... (post-pre) comment (a) Genome Biology 2007, Imiquimod Vehicle Before treatment (Px-PB1) q12,2D q12,4D ii iii q24,8D iv 23 genes 65 genes 263 56 genes 200 23 65 18 v refereed research 23 deposited research...

Ngày tải lên: 14/08/2014, 17:22

... biases when searching for horizontally transferred genes Mol Biol Evol 2001, 18:1838-1840 Tsirigos A, Rigoutsos I: A new computational method for the detection of horizontal gene transfer events Nucleic ... links phylogenetic information for thousands of different species to millions of protein sequences One notable exception has been the work of Koonin et al [1], who searched for horizontal transfer ... horizontal gene transfer PLoS Biol 2005, 3:e316 Gophna U, Charlebois RL, Doolittle WF: Have archaeal genes contributed to bacterial virulence? Trends Microbiol 2004, 12:213-219 Faguy DM: Lateral gene transfer...

Ngày tải lên: 14/08/2014, 17:22

... virus-1 plasmid vectors for gene therapy of Parkinson's disease and other neurological disorders Exp Neurol 1997, 144:98-102 Gentry T, Smith C: Retroviral vector- mediated gene transfer into umbilical ... advantage for the system is that vector DNA could possibly be maintained as an episome with minimal concern for the potential consequences of random integration of vector DNA (i.e activation of oncogenes ... cells Advantages for use of the HCMV amplicon Foremost among the advantages of the vector system we have described is the potential ability to efficiently infect and deliver genetic information to...

Ngày tải lên: 14/08/2014, 19:22

... performed to verify the results For the luciferase assay, three mice were set up per time point /gene transfer vector treatment and one mouse was used as control for each time point/control vector ... Intranasal inoculation of adenoviral gene transfer vectors For whole organ GFP imaging analysis, a replicationdefective recombinant human type adenoviral vector expressing green fluorescent protein ... recombinant adenoviral gene transfer vector, we first used a recombinant replication-deficient adenoviral vector expressing green fluorescent protein (AdGFP) Use of this vector allowed us to conveniently...

Ngày tải lên: 14/08/2014, 19:22

... Squadrito F: Adeno-associated viral vector- mediated human vascular endothelial growth factor gene transfer stimulates angiogenesis and wound healing in the genetically diabetic mouse Diabetologia ... regression of vascular structures Therefore, the lack of effect of AAV-FGF4-IRES-VEGF-A vector on the total number of skin capillaries and their proliferation 21 days after wounding and gene transfer ... Page of 16 Figure In vitro gene expression in AAV-transduced HeLa cells (A) Schematic representation of expression cassettes in AAV vectors used for transduction: control vector encoding b-galactosidase...

Ngày tải lên: 14/08/2014, 19:22

... heterogenicity of vtg gene family and expressions of multiple vtg genes were investigated using a model fish, the zebrafish (Danio rerio) The study of vtg genes provides valuable information for further ... Chapter General conclusions Based on the findings presented in this thesis, the following general conclusions can be drawn: 1) At least seven distinct vitellogenin genes (vtgs) were ... the traditional belief that liver is the only organ for Vtg synthesis in oviparous vertebrates The preliminary studies on receptor-mediated gene transfer using fish Vtgs as DNA carriers encourage...

Ngày tải lên: 16/09/2015, 17:13

... weight was 29.1 kDa for 6xHis-LVIa, 57.3 kDa for 6xHis-LVIb, 39 kDa for 6xHis-LVIc, 45.4 kDa for 6xHis-LVII, 27.7 kDa for 6xHis-LVIb1, 25.1 kDa for 6xHis-LVIb2 and 24.3 kDa for 6xHis-LVIb3 Protein ... mRNA/cDNA were performed at 94°C for min, followed by 35 to 40 cycles of PCR for amplification of target sequences using the following conditions: 94 °C for 30 sec, 55~67 °C for min, 72 °C for min, final ... 95 ºC for prior to loading (for SDS-PAGE) or directly loaded without denaturation (for NPAGE) Gel electrophoresis was performed with constant voltage at 50 V for 0.5 h, followed by 100 V for h...

Ngày tải lên: 16/09/2015, 17:13

... performed at 55ºC for 10 min, followed by denaturation at 95ºC for 30 s Subsequent PCR were performed for 45 cycles with the parameters for each cycle as following: denaturation at 95 ºC for ... place at ºC for later use 3.2.5.2 Tissue preparation before hybridization Tissue sections were de-waxed in two changes of Histo-Clear II for 10 each and rinsed in 100% ethanol for before re-hydration ... pBluescrip SK vector, 20-bp downstream of each insert’s poly-A tail (see Figs 2-1, 2-4 to 2-7 in Chapter 2; vector map not shown) Probes for vtg2, vtg3 and β-actin were generated by PCR Briefly, a gene...

Ngày tải lên: 16/09/2015, 17:13

... DNA The reaction was performed for 25 cycles with the following parameters for each cycle: denaturing at 96 ºC for 10 sec, annealing at 50 ºC for sec and extension at 60 ºC for The sequencing reaction ... PCR was performed on a DNA Thermal Cycler 480 (Perkin Elmer) with the following parameters: 94 oC for min; 35 cycles of 94 oC for 30 sec, 60 oC for and 72 oC for min; finally 72 oC for A PCR fragment ... described for the 5’ RACE PCR except for the template PCR was also performed on the DNA Thermal Cycler 480 with the following parameters: 94 oC for min; 30 cycles of 94 oC for 30 sec, 54 oC for and...

Ngày tải lên: 16/09/2015, 17:13

... technology Thus, gene transfer, as the first step in making transgenic fish, is obviously very important The techniques of gene transfer vary and have progressively improved 1.3.2 Current gene transfer ... transgene on to their offspring) (Hostetler et al., 2003) This approach seems promising in the future C Sperm-mediated gene transfer The first attempt using sperm as vectors for introducing foreign ... 1993) The DNA transfer rate in this case is not very high Although sperm-mediated gene transfer has been successfully used to generate transgenic fish, full harness of this gene transfer method...

Ngày tải lên: 16/09/2015, 17:13

... forms of the approach, such as transgenICSI However, AI-based SMGT is not yet an established method of transgenesis, therefore the prospects for this potentially very important form of gene transfer ... established for humans, thus ESC-based gene transfer remains – despite its effectiveness in mice – unavailable for human germline gene therapy The lack of human ESCs leaves NT-based gene transfer ... sperm-mediated gene transfer Transplant Proc 1999; 31(1-2):972-974 42 Lazzereschi D, Forni M, Cappello F, et al Efficiency of transgenesis using sperm-mediated gene transfer: Generation of hDAF...

Ngày tải lên: 03/11/2012, 10:01

... in the city, who might not be representative for Vietnamese middle class in other areas, and difenitely not valid for average Vietnamese shoppers in general However, they can represent a segment ... business formula used in this industry There are criteria used to define a supermarket: the minimum size requirements, the characteristics and extent of self-service, and the stock requirements ... Foreign investment was not allowed until recently and even now foreign investment is limited to 28 stores not exceeding 800 sq m Existing retailers are sophisticated , aggressive and big In general...

Ngày tải lên: 13/04/2013, 10:30

... profile Deformed reinforcement — Deformed reinforcing bars, bar mats, deformed wire, welded plain wire fabric, and welded deformed wire fabric conforming to 3.5.3 Deformed reinforcement — Deformed ... only for spiral reinforcement (either as lateral reinforcement for compression members, for torsion members, or for confining reinforcement for splices) 3.5.4.2 — Plain wire for spiral reinforcement ... mats for concrete reinforcement shall conform to “Specification for Fabricated Deformed Steel Bar Mats for Concrete Reinforcement” (ASTM A 184) Reinforcing bars used in bar mats shall conform...

Ngày tải lên: 03/08/2013, 11:24

... load a driver for a function not currently attached to the bus Figure Generic bus driver model when two functions are contained in one device Bus Driver and Device Requirements for a Generic Driver ... installed on the platform Problems with this method are: • Detection methods used by drivers might conflict with hardware already installed on the platform For example, if a platform has a drill ... meet the requirements of discoverability and multiplexability • Discoverable - If special information is required in descriptors to group interfaces into functions, this is a problem for a generic...

Ngày tải lên: 07/10/2013, 00:20