Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: DrugBank:EXPT01586 (G418)
 2,237 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

To study and evaluate the potential of the haematopoietic system as a target for gene therapy in haemophilia A, we have infected murine bone-marrow cells with a recombinant retrovirus encoding blood-coagulation factor VIII and the bacterial enzyme neomycin-phosphotransferase. After transplantation of the infected bone marrow into lethally irradiated mice, the presence of intact vector could be demonstrated in DNA isolated from individual haematopoietic progenitor-cell-derived spleen colonies. About 8% of the spleen colonies were shown to contain the intact vector. Selection for resistance to the neomycin analogue G418 prior to transplantation specifically killed the uninfected bone-marrow cells and, as a result, over 90% of the spleen colonies contained the factor VIII vector. However, expression of factor VIII in vivo, either at the RNA or at the protein level could not be demonstrated. From these data we conclude that: 1) retroviral vectors can be used to transfer factor-VIII cDNA into haematopoietic progenitor cells; 2) the vector sequences are expressed immediately after integration; and 3) transcription of the vector is repressed in the progenitor-cell-derived cells.
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PMID:Toward gene therapy in haemophilia A: retrovirus-mediated transfer of a factor VIII gene into murine haematopoietic progenitor cells. 164 25

A retrovirus-mediated transduction of B-domain-deleted human blood coagulation factor VIII (FVIII-B) was attempted in partially hepatectomized rats. FVIII-B cDNA was inserted into a retroviral vector (pLNSX) and infective recombinant virus particles were produced in packaging cell lines (psi2 and PA317). Transfection of mouse NIH-3T3 cells with the FVIII-B cDNA inserted recombinant viruses, followed by G418 selection, gave a viral titer of 3.5 x 10(4) CFU/ml. FVIII-B protein, as well as FVIII-B mRNA, was detected in these cells. Transfusion of FVIII-B-expressing retrovirus particles into the tail vein of rats subjected to partial hepatectomy resulted in a relatively higher level of FVIII-B expression in liver and circulating plasma as compared with the sham-operated rats. These results indicate that the augmentation of FVIII activity in the blood of an animal by retroviral gene delivery can be enhanced by partial hepatectomy, and that the retrovirus-mediated FVIII-B cDNA delivery to regenerating liver may be an alternative method for the expression of FVIII-B cDNA in vivo.
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PMID:An efficient retrovirus-mediated transduction of human blood coagulation factor VIII cDNA in regenerating rat liver. 1039 Nov 1

To investigate the non-viral vector mediating human coagulation factor VIII gene expression in mouse 32D cell line, a recombinant plasmid vector, pRC/RSV-hFVIIIBDcDNA, was constructed by cloning B-domain-deleted (Delta760aa-1639aa) human factor VIII cDNA (hFVIIIBDcDNA) into plasmid vector, pRC/RSV. The plasmid RC/RSV-hFVIIIBDcDNA was then transfected by means of SuperFect Transfection Reagent into mouse 32D cell line. After screening with G418, the procoagulant activity (hFVIII:C) and antigen (hFVIII:Ag) of human factor VIII in the culture medium were detected using one-stage method and ELISA, respectively. Furthermore, RT-PCR was performed to observe the transcription of hFVIIIBDcDNA. The results showed that human coagulation factor VIII protein existed in culture medium with hFVIII:C up to 2.01 U/(10(6) cell x 24 hours) and hFVIII:Ag to 450.08 ng/(10(6) cell x 24 hours). RT-PCR displayed mRNA of hFVIIIBDcDNA in 32D cells. It is concluded that the recombinant plasmid RC/RSV-hFVIIIBDcDNA can successfully express human FVIII in mouse 32D cell line, and hFVIII expressed in vitro presents the similar coagulant activity to the native hFVIII existing in normal human plasma.
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PMID:[Non-viral vector mediating human coagulation factor VIII gene expression in mouse 32D cell line]. 1563 47