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1.
Delivery of DNA into mammalian cells by receptor-mediated endocytosis and gene therapy 总被引:8,自引:0,他引:8
The correction of genetically based disorders by the introduction of a therapeutic genetic construct into the appropriate
cell type (“gene therapy”), has become a distinct possibility in recent years. In order for gene therapy to be a practical
alternative to more conventional pharmaceutical approaches to treatment, it must be administrable in vivo. This demands that
a system be developed that can specifically target the DNA to the desired cell type once introduced into the patient. Among
the procedures that are currently being pursued, the delivery of DNA to cells by receptor mediated endocytosis (RME), comes
closest to fulfilling this crucial requirement.
The natural physiological process of RME can be exploited to deliver genetic material to cells. An antibody or ligand to a
cell surface receptor that is known to undergo endocytosis, is complexed with DNA through a covalently linked polycationic
adjunct (e.g., polylysine, protamines). Such complexes retain their binding specificity to the cell surface and are taken
up into the cell where they enter the endosomal compartment via normal endocytotic processes. In addition, steps must be taken
to avoid degradation of the DNA within the endosome-lysosome. Cells can be treated with the lysosomatropic agent chloroquine
during the transfection procedure. Alternatively, the components of viruses that enter cells by endocysis and possess an endosomal
“break out” capacity can be used. Replication defective adenovirus coupled to the ligand-DNA complex gives transfection efficiencies
of virtually 100% on tissue culture cells in vitro. Synthetic peptides that mimic the membrane fusing region of influenza
virus hemagglutinin, have also been successfully used as part of the ligand-DNA complex to bring about endosomal escape.
Preliminary studies have demonstrated the potential of this method to specifically target DNA to the cell type of choice in
vivo. Delivery of genes by receptor-mediated endocytosis offers the greatest hope that gene therapy can be an inexpensive,
easily applicable, widespread technology. 相似文献
2.
M. Pesic K. Drabek C. Esler S. Ruzdijic V. Pejanovic Z. Pietrzkowski 《Nucleosides, nucleotides & nucleic acids》2013,32(5-6):963-975
Abstract 8–Cl-cAMP and tiazofurin (TR) are anti-tumor agents that besides their antiproliferative effect, also induce differentiation of tumor cells. Although, these agents exert a profound effect on the same events of tumor cell life, it is thought that 8-Cl-cAMP and TR act by modulating the signal transduction pathway through distinct mechanisms. We have compared their effect on two human glioma cell lines (U87 MG and U251 MG) and examined if there is selectivity in their action toward normal human astrocytes. 相似文献
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Pesic M Drabek K Esler C Ruzdijic S Pejanovic V Pietrzkowski Z 《Nucleosides, nucleotides & nucleic acids》2000,19(5-6):963-975
8-Cl-cAMP and tiazofurin (TR) are anti-tumor agents that besides their antiproliferative effect, also induce differentiation of tumor cells. Although, these agents exert a profound effect on the same events of tumor cell life, it is thought that 8-Cl-cAMP and TR act by modulating the signal transduction pathway through distinct mechanisms. We have compared their effect on two human glioma cell lines (U87 MG and U251 MG) and examined if there is selectivity in their action toward normal human astrocytes. 相似文献
6.
Developments in insect transgenesis using transposons combined with available mass rearing technology for insects such as
the Medfly, Ceratitis capitata, provide opportunity for the production of protein for industrial, agricultural and healthcare purposes on a very large scale.
In this study, we report the germ-line transformation and expression of a cDNA encoding human growth hormone (hGH) in transgenic
Drosophila using the Minos transposon. Production and secretion of a bioactive hGH into the haemolymph of transgenic larvae was demonstrated by immunoblot
analysis, ELISA and a proliferation bioassay. Stable expression of hGH was observed over 50 generations. The results indicate
that mass reared transgenic diptera with a rapid period of larval growth could provide cost effective production systems for
the manufacture of therapeutic and other high value proteins. 相似文献
7.
ML Zupancic BL Cantarel Z Liu EF Drabek KA Ryan S Cirimotich C Jones R Knight WA Walters D Knights EF Mongodin RB Horenstein BD Mitchell N Steinle S Snitker AR Shuldiner CM Fraser 《PloS one》2012,7(8):e43052
Obesity has been linked to the human gut microbiota; however, the contribution of gut bacterial species to the obese phenotype remains controversial because of conflicting results from studies in different populations. To explore the possible dysbiosis of gut microbiota in obesity and its metabolic complications, we studied men and women over a range of body mass indices from the Old Order Amish sect, a culturally homogeneous Caucasian population of Central European ancestry. We characterized the gut microbiota in 310 subjects by deep pyrosequencing of bar-coded PCR amplicons from the V1-V3 region of the 16S rRNA gene. Three communities of interacting bacteria were identified in the gut microbiota, analogous to previously identified gut enterotypes. Neither BMI nor any metabolic syndrome trait was associated with a particular gut community. Network analysis identified twenty-two bacterial species and four OTUs that were either positively or inversely correlated with metabolic syndrome traits, suggesting that certain members of the gut microbiota may play a role in these metabolic derangements. 相似文献
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Charlotte Krahé Marianne M. Drabek Yannis Paloyelis Aikaterini Fotopoulou 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》2016,371(1708)
Affective touch and cutaneous pain are two sub-modalities of interoception with contrasting affective qualities (pleasantness/unpleasantness) and social meanings (care/harm), yet their direct relationship has not been investigated. In 50 women, taking into account individual attachment styles, we assessed the role of affective touch and particularly the contribution of the C tactile (CT) system in subjective and electrophysiological responses to noxious skin stimulation, namely N1 and N2-P2 laser-evoked potentials. When pleasant, slow (versus fast) velocity touch was administered to the (non-CT-containing) palm of the hand, higher attachment anxiety predicted increased subjective pain ratings, in the same direction as changes in N2 amplitude. By contrast, when pleasant touch was administered to CT-containing skin of the arm, higher attachment anxiety predicted attenuated N1 and N2 amplitudes. Higher attachment avoidance predicted opposite results. Thus, CT-based affective touch can modulate pain in early and late processing stages (N1 and N2 components), with the direction of effects depending on attachment style. Affective touch not involving the CT system seems to affect predominately the conscious perception of pain, possibly reflecting socio-cognitive factors further up the neurocognitive hierarchy. Affective touch may thus convey information about available social resources and gate pain responses depending on individual expectations of social support.This article is part of the themed issue ‘Interoception beyond homeostasis: affect, cognition and mental health’. 相似文献
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Drabek K van Ham M Stepanova T Draegestein K van Horssen R Sayas CL Akhmanova A Ten Hagen T Smits R Fodde R Grosveld F Galjart N 《Current biology : CB》2006,16(22):2259-2264
In motile fibroblasts, stable microtubules (MTs) are oriented toward the leading edge of cells. How these polarized MT arrays are established and maintained, and the cellular processes they control, have been the subject of many investigations. Several MT "plus-end-tracking proteins," or +TIPs, have been proposed to regulate selective MT stabilization, including the CLASPs, a complex of CLIP-170, IQGAP1, activated Cdc42 or Rac1, a complex of APC, EB1, and mDia1, and the actin-MT crosslinking factor ACF7. By using mouse embryonic fibroblasts (MEFs) in a wound-healing assay, we show here that CLASP2 is required for the formation of a stable, polarized MT array but that CLIP-170 and an APC-EB1 interaction are not essential. Persistent motility is also hampered in CLASP2-deficient MEFs. We find that ACF7 regulates cortical CLASP localization in HeLa cells, indicating it acts upstream of CLASP2. Fluorescence-based approaches show that GFP-CLASP2 is immobilized in a bimodal manner in regions near cell edges. Our results suggest that the regional immobilization of CLASP2 allows MT stabilization and promotes directionally persistent motility in fibroblasts. 相似文献
10.
Grinblat-Huse V Drabek EF Creasy HH Daugherty SC Jones KM Santana-Cruz I Tallon LJ Read TD Hatch TP Bavoil P Myers GS 《Journal of bacteriology》2011,193(15):4039-4040
Chlamydia psittaci is a highly prevalent avian pathogen and the cause of a potentially lethal zoonosis, causing life-threatening pneumonia in humans. We report the genome sequences of C. psittaci 6BC, the prototype strain of the species, and C. psittaci Cal10, a widely used laboratory strain. 相似文献