fetal muscle gene therapy gene delivery in large animals

... transcriptsin both slow twitch skeletal muscle as well as the heart It binds Ca2+ and prevents actin-myosin interaction in resting muscle Page of (page number not for citation purposes) Genetic Vaccines ... factors) binding sites as well as NFkappaB binding sites The CMV promoter confers virtually ubiquitous expression throughout the body except in the liver where it becomes inactive once the initial inflammatory ... expression were observed in hearts of animals injected with the CMV, Des or α-MHC constructs (Figure 1B) Expression levels in hearts of animals injected with viruses containing either the MLC-2 or...

Ngày tải lên: 14/08/2014, 19:22

... SMC, by inhibiting signalling molecules [45–47] Caveolin-3 is the main isoform in muscle cells, including SMC [30] Caveolin-1 and caveolin-3 seem to act as scaffolding proteins that negatively regulate ... MAPK kinase (MEK) inhibitor, U0126 U0126 completely inhibited the cyclin D1 promoter activity in AA-pretreated SMC incubated with IL-1b and reduced it in cells incubated with fetal bovine serum ... stimulated (up to threefold) by incubation in medium containing fetal bovine serum In parallel to their effects on the cyclin D1 gene promoter, EPA and DHA also reduced the egr-1 gene promoter activity...

Ngày tải lên: 19/02/2014, 16:20

... protein kinase kinase (MEK) inhibitor, U0126 [32,33] Therefore, the role of the MAPK pathway in regulating PDK mRNA expression was investigated by incubating myoblasts for h in the presence of insulin ... used to examine the role of PtdIns3K in regulating PDK transcript abundance in response to insulin (Fig 2) Incubation with either LY294002 (50 lm) or wortmannin (100 nm) significantly inhibited ... mammalian target of rapamycin (mTOR) GSK3 is inactivated in response to insulin via a PtdIns3K ⁄ protein kinase B (PKB)dependent pathway [27,28] Involvement of GSK3 in FEBS Journal 272 (2005)...

Ngày tải lên: 07/03/2014, 17:20

... cytofectins for gene delivery M Spelios and M Savva Cationic lipidpDNA binding studies Binding curves of EtBr-intercalated pDNA titrated with aliquots of cationic lipid dispersions are shown in Fig ... synthesized with the intent of enhancing the transport of exogenous genetic material into cells in vitro, and their design eliminates the need for additional components in the gene- delivery system ... and maintained in serum medium (Dulbeccos modied Eagles medium supplemented with 10% fetal bovine serum, 50 unitsặmL)1 penicillin and 50 lgặmL)1 streptomycin) After incubation for h at 37 C in a...

Ngày tải lên: 23/03/2014, 07:20

... assays (EMSAs) using radiolabeled oligonucleotides containing either a STAT5B-binding site or an AP-1-binding site in the rat AGT gene promoter As shown in Fig 2A,B, DNA–protein complexes were ... c-Jun may be involved in Ang II-induced AGT mRNA expression (Fig 1C) DNA-binding activity of STAT5B and c-Jun increases in Ang II-induced AGT expression To find out whether the increase in STAT5B ... oligonucleotide probes containing the AP-1-binding site (A) and the STAT5B-binding site (B) in the AGT gene promoter Protein–DNA complexes were separated by nondenaturing PAGE and then visualized...

Ngày tải lên: 30/03/2014, 02:20

... Douillard-Guilloux, and Catherine Caillaud Part III Methods for Muscle Gene Transfer in Large Animal Models 15 Fetal Muscle Gene Therapy /Gene Delivery in Large Animals Khalil ... With this in mind, we compiled this collection of muscle gene therapy methods and protocols with the intention of bridging the translational gap in muscle gene therapy The book is divided into three ... enhancing activity of the cassette and staying within the vector packaging size constraints (e.g., introducing a large insertion into a cassette may not be justified by only a slight increase in...

Ngày tải lên: 29/06/2014, 09:20

... Zaboikin M, Srinivasakumar N, Zaboikina T, Schuening F: Cloning and expression of canine O6-methylguanine-DNA methyltransferase in target cells, using gammaretroviral and lentiviral vectors Hum Gene ... Since SIV RRE had not been tested in an HIV-1 packaging system, several packaging and gene transfer vectors containing HIV-1 or SIV RREs were created (Figure 1) The packaging constructs contained ... titers were obtained in the presence of HIV-1 Rev or HTLV-1 Rex The SIV Rev achieved only a marginal increase in titer over that obtained in the absence of any 'transport' protein expression construct...

Ngày tải lên: 10/08/2014, 05:21

... a-terpineol; (B) Pg×eTPS-Cin: (+)-a-pinene, b-pinene, myrcene, 1,8-cineole, g-terpinene, unknown, (-)-a-terpineol, unknown; (C) PgTPS-Lin: (-)-linalool; (D) PsTPS-Phel-1: (-)-a-pinene, (-)-bpinene, ... Others WS0291_H24 4.7 b-Pinene PsTPS-Cin 1.1 89.1 (+)-a-Pinene WS00921_D15 0.8 Others 1,8-Cineole (-)-a-Terpineol Pg×eTPS-Cin 1.4 g-Terpinene WS02628_N22 2.1 a-Terpineol PgTPS-Cin 2.1 Myrcene Functional ... synthases: (-)-a/b-pinene synthases We characterized one new (-)-a/b-pinene synthase in Sitka spruce (PsTPS-Pin) and two in white spruce (PgTPS-Pin-1 and PgTPS-Pin-2; both originating from the same...

Ngày tải lên: 11/08/2014, 11:21

... to the reduction in numbers of lipid droplets, the layer of basal infoldings was also thinner in rAAV.RPE65-injected eyes (Fig 6A and 6B) Immunostaining using the anti-SWC opsin antibody demonstrated ... encoding Prph2 in a retinal degeneration slow mouse model [42,43] Vision is maintained through the close and precise interaction between all the retinal cells For example both in humans and in ... immunohistochemical labelling in the retinas of Rpe65-/- mice after injection with rAAV.RPE65 Labeling in the retinal pigment epithelium is seen at mo post-injection (A) The signal continues for some distance...

Ngày tải lên: 14/08/2014, 19:22

... necessity? Insertional mutagenesis in gene therapy and its consequences Mol Ther 2004, 9:5-13 Dave UP, Jenkins NA, Copeland NG: Gene therapy insertional mutagenesis insights Science 2004, 303:333 Hacein-Bey ... transduction by using multiple cytokines to stimulate division of PHSC This is likely to induce many genes involved in regulating cell growth As retroviruses preferentially integrate into active gene sequences, ... is a gene capable of influencing cell growth (although in neither case can it be considered a classical oncogene) In terms of the influence of vector design it is interesting to note that in both...

Ngày tải lên: 14/08/2014, 19:22

... Genetic Vaccines and Therapy 2004, 2:16 Introduction In vivo gene targeting of highly specific cell subsets remains the main challenge for gene therapy of a broad range ... administered conjugates strongly determine their in vivo performance These include avid interaction with serum components, resulting in colloidal instability, including both aggregation and dissociation ... recently, in vivo, we have shown a specific tumor targeting after a single intravenous injection in mice bearing tumour expressing the renal carcinoma – related G250 tumor associated antigen [16]...

Ngày tải lên: 14/08/2014, 19:22

... an emerging vector 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 ... continuously in a real-time manner in individual animals thus reducing the subject-to-subject variation and minimizing the number of animals needed in the test; and the commercial available imaging ... 37ºC in a humidified incubator with 100% air Insect Sf9 cell line purchased from Invitrogen was cultured in Sf-900 II SFM medium with penicillin streptomycin (0.5%) at 27ºC in a non-humidified incubator...

Ngày tải lên: 01/10/2015, 11:39

... of immunoglobulin gene, γIg – the γ-chain of immunoglobulin gene, IFN-ind – the interferone-inducible gene 6-16, IL4R – the interleukin receptor gene, TAP2 – the transport protein gene, hnRNP A1 ... Hodgkin’s disease; capn4: the calpain subunit gene; ND4: the NADH dehydrogenase subunit gene; IL5: the interleukin gene; λIg: the λ-chain of immunoglobulin gene; γIg: the γ-chain of immunoglobulin ... immunoglobulin gene; IFN-ind: the interferon-inducible gene 6-16; IL4R: the interleukin receptor gene; TAP2: the transport protein gene; hnRNP A1: the human ribonucleoprotein А1 gene; SMG-1 :...

Ngày tải lên: 02/11/2012, 11:08

... the genetic consequences of targeted gene therapy in vitro (and in vivo) Microarray Study Differential Gene Expression Differential gene expression profiling is routinely used to detect genes ... protein kinase N1 is a transcript variant encoded by PKN Its cytoplasmic gene product is a serine/threonine-protein kinase N1 which is a member of the protein kinase C superfamily (http://amigo.geneontology.org/cgi-bin/amigo/ ... polymer); the ordinates represent the linear ratio of differential gene expression Genes detected and in following investigated for validation A) an induced expression of the PKN1 gene B) a repressed...

Ngày tải lên: 03/11/2012, 11:44

... of cyclin protein levels or by inducing members of the INK4A and/or CIP/KIP family of cyclin dependent kinase inhibitors (CKI), resulting in the down-regulation of cyclin/Cdk activity and inhibition ... Berns A (2002) Tumor formation in mice with somatic inactivation of the retinoblastoma gene in interphotoreceptor retinal binding protein-expressing cells Oncogene 21:4635–4645 Voorhoeve PM, ... while the pocket-domain and the C-domain are involved in binding E2F proteins Mdm2 (as well as c-Abl) binds to the C-domain All pocket proteins are subject to phosphorylation (indicated with “P”)...

Ngày tải lên: 25/10/2013, 21:20

... McClain DA, Litwin SE et al (2002) Insulin signaling coordinately FEBS Journal 274 (2007) 5278–5284 ª 2007 The Authors Journal compilation ª 2007 FEBS 5283 Metabolic gene switching in murine female ... substrate In agreement, lower PDK-4 gene expression in female hearts indicates that 5280 myocardial glucose metabolism may be increased in parallel As optimization of glucose metabolism is increasingly ... metabolic gene switch (decrease in fatty acid utilization, increase in glucose utilization) in the murine female heart Moreover, our data suggest that these changes occur in an Akt-independent...

Ngày tải lên: 18/02/2014, 16:20

... functional guinea pig genes Interestingly, we observed the greatest differences between these closely related genes within intron and intron This could indicate frequent recombination between the genes ... for an altered binding site for adrenodoxin or modified binding affinities Also, experiments in a reconstituted system with bovine CYP11B0 using mutant forms of adrenodoxin that have increased electron ... proteins of the CYP11B family including CYP11B2 of the guinea pig have a basic residue (arginine or lysine, respectively) at the corresponding position These two basic residues have been shown in...

Ngày tải lên: 22/02/2014, 07:20

... examined in mice As shown in Figure 3, PM 2.5 exposure induced a marked increase in macrophage (F4/80+ cells) infiltration in eWAT Next, we analyzed the changes in uncoupling protein (UCP1) in ... long chain fatty acids (Elovl3), type iodothyronine deiondinase (Dio2), homeobox C9 (Hoxc9), insulin-like growth factor binding protein (Igfbp3), dermatopontin (Dpt), and b-actin are showed in Table ... Immunohistochemical examination of uncoupling protein (UCP1) in the iBAT A iBAT was stained by antibody against UCP1 and counterstained with hematoxylin B Quantification of UCP1 in iBAT n = *P < 0.05...

Ngày tải lên: 06/03/2014, 19:20

... 15 kDa protein, ileal bile acid-binding protein and gastrotropin, reflecting the speculations of authors on its intracellular function(s) In vitro binding assays revealed a surprisingly low affinity ... Intracellular lipid-binding proteins and their genes Annu Rev Nutr 17, 277–303 Schaap FG, Van der Vusse GJ & Glatz JFC (2002) Evolution of the family of intracellular lipid binding proteins in ... identified in vertebrate species [3] Originally, these proteins were named according to the tissue from which they were initially isolated, e.g livertype fatty acid-binding protein (L-FABP), brain-type...

Ngày tải lên: 07/03/2014, 06:20

... Thirteen genes, which encode six protein families responsible for protein folding, are included in the UPR gene list Among them, BiP (three genes), calnexin (two genes), calreticulin (two genes), ... ERSE-like Xbp1 binding-like P-UPRE ERSE-like ERSE-like ERSE-like Xbp1 binding-like Xbp1 binding-like ERSE-like – ERSE-like Xbp1 binding-like Xbp1 binding-like ERSE-like – ERSE-like – Xbp1 binding-like ... equilibrated with the urea-binding buffer After washing the cartridge with the urea-binding buffer, the luminal domain of At SEL-1L was eluted with the urea-binding buffer containing m imidazole, and...

Ngày tải lên: 07/03/2014, 21:20