scholarly journals Replication-defective chimeric helper proviruses and factors affecting generation of competent virus: expression of Moloney murine leukemia virus structural genes via the metallothionein promoter.

1987 ◽  
Vol 7 (5) ◽  
pp. 1797-1806 ◽  
Author(s):  
R A Bosselman ◽  
R Y Hsu ◽  
J Bruszewski ◽  
S Hu ◽  
F Martin ◽  
...  
Keyword(s):  
Cell Line ◽  
Sequence Homology ◽  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Moloney Murine Leukemia Virus ◽  
Helper Cell ◽  
Factors Affecting ◽  
Cis Acting ◽  
Virus Expression ◽  
Murine Leukemia

Two chimeric helper proviruses were derived from the provirus of the ecotropic Moloney murine leukemia virus by replacing the 5'long terminal repeat and adjacent proviral sequences with the mouse metallothionein I promoter. One of these chimeric proviruses was designed to express the gag-pol genes of the virus, whereas the other was designed to express only the env gene. When transfected into NIH 3T3 cells, these helper proviruses failed to generate competent virus but did express Zn2+-inducible trans-acting viral functions needed to assemble infectious vectors. One helper cell line (clone 32) supported vector assembly at levels comparable to those supported by the Psi-2 and PA317 cell lines transfected with the same vector. Defective proviruses which carry the neomycin phosphotransferase gene and which lack overlapping sequence homology with the 5' end of the chimeric helper proviruses could be transfected into the helper cell line without generation of replication-competent virus. Mass cultures of transfected helper cells produced titers of about 10(4) G418r CFU/ml, whereas individual clones produced titers between 0 and 2.6 X 10(4) CFU/ml. In contrast, defective proviruses which share homologous overlapping viral sequences with the 5' end of the chimeric helper proviruses readily generated infectious virus when transfected into the helper cell line. The deletion of multiple cis-acting functions from the helper provirus and elimination of sequence homology overlapping at the 5' ends of helper and vector proviruses both contribute to the increased genetic stability of this system.

Replication-defective chimeric helper proviruses and factors affecting generation of competent virus: expression of Moloney murine leukemia virus structural genes via the metallothionein promoter

1987 ◽  
Vol 7 (5) ◽  
pp. 1797-1806
Author(s):  
R A Bosselman ◽  
R Y Hsu ◽  
J Bruszewski ◽  
S Hu ◽  
F Martin ◽  
...  
Keyword(s):  
Cell Line ◽  
Sequence Homology ◽  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Moloney Murine Leukemia Virus ◽  
Helper Cell ◽  
Factors Affecting ◽  
Cis Acting ◽  
Virus Expression ◽  
Murine Leukemia

Two chimeric helper proviruses were derived from the provirus of the ecotropic Moloney murine leukemia virus by replacing the 5'long terminal repeat and adjacent proviral sequences with the mouse metallothionein I promoter. One of these chimeric proviruses was designed to express the gag-pol genes of the virus, whereas the other was designed to express only the env gene. When transfected into NIH 3T3 cells, these helper proviruses failed to generate competent virus but did express Zn2+-inducible trans-acting viral functions needed to assemble infectious vectors. One helper cell line (clone 32) supported vector assembly at levels comparable to those supported by the Psi-2 and PA317 cell lines transfected with the same vector. Defective proviruses which carry the neomycin phosphotransferase gene and which lack overlapping sequence homology with the 5' end of the chimeric helper proviruses could be transfected into the helper cell line without generation of replication-competent virus. Mass cultures of transfected helper cells produced titers of about 10(4) G418r CFU/ml, whereas individual clones produced titers between 0 and 2.6 X 10(4) CFU/ml. In contrast, defective proviruses which share homologous overlapping viral sequences with the 5' end of the chimeric helper proviruses readily generated infectious virus when transfected into the helper cell line. The deletion of multiple cis-acting functions from the helper provirus and elimination of sequence homology overlapping at the 5' ends of helper and vector proviruses both contribute to the increased genetic stability of this system.

scholarly journals CrFK Feline Kidney Cells Produce an RD114-Like Endogenous Virus That Can Package Murine Leukemia Virus-Based Vectors

1998 ◽  
Vol 72 (9) ◽  
pp. 7685-7687 ◽  
Author(s):  
Jörg G. Baumann ◽  
Walter H. Günzburg ◽  
Brian Salmons
Keyword(s):  
Cell Line ◽  
Kidney Cell ◽  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Moloney Murine Leukemia Virus ◽  
Viral Infectivity ◽  
Kidney Cells ◽  
Kidney Cell Line ◽  
Endogenous Virus ◽  
Murine Leukemia

ABSTRACT The feline kidney cell line CrFK is used extensively for viral infectivity assays and for study of the biology of various retroviruses and derived vectors. We demonstrate the production of an endogenous, RD114-like, infectious retrovirus from CrFK cells. This virus also is shown to efficiently package Moloney murine leukemia virus vectors.

scholarly journals A Chimeric Ty3/Moloney Murine Leukemia Virus Integrase Protein Is Active In Vivo

1998 ◽  
Vol 72 (5) ◽  
pp. 4297-4307 ◽  
Author(s):  
Sandra L. Dildine ◽  
James Respess ◽  
Doug Jolly ◽  
Suzanne B. Sandmeyer
Keyword(s):  
Cell Line ◽  
Murine Leukemia Virus ◽  
Genomic Dna ◽  
Integration Site ◽  
Leukemia Virus ◽  
Moloney Murine Leukemia Virus ◽  
Packaging Cell Line ◽  
Terminal Domain ◽  
Murine Leukemia

ABSTRACT This report describes the results of experiments to determine whether chimeras between a retrovirus and portions of Ty3 are active in vivo. A chimera between Ty3 and a Neor-marked Moloney murine leukemia virus (M-MuLV) was constructed. The C-terminal domain of M-MuLV integrase (IN) was replaced with the C-terminal domain of Ty3 IN. The chimeric retroviruses were expressed from an amphotrophic envelope packaging cell line. The virus generated was used to infect the human fibrosarcoma cell line HT1080, and cells in which integration had occurred were selected by G418 resistance. Three independently integrated viruses were rescued. In each case, the C-terminal Ty3 IN sequences were maintained and short direct repeats of the genomic DNA flanked the integration site. Sequence analysis of the genomic DNA flanking the insertion did not identify a tRNA gene; therefore, these integration events did not have Ty3 position specificity. This study showed that IN sequences from the yeast retrovirus-like element Ty3 can substitute for M-MuLV IN sequences in the C-terminal domain and contribute to IN function in vivo. It is also one of the first in vivo demonstrations of activity of a retrovirus encoding an integrase chimera. Studies of chimeras between IN species with distinctive integration patterns should complement previous work by expanding our understanding of the roles of nonconserved domains.

Hypoxia inhibits Moloney murine leukemia virus expression in activated macrophages

2006 ◽  
Vol 81 (2) ◽  
pp. 528-538 ◽  
Author(s):  
Maura Puppo ◽  
Maria Carla Bosco ◽  
Maurizio Federico ◽  
Sandra Pastorino ◽  
Luigi Varesio
Keyword(s):  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Moloney Murine Leukemia Virus ◽  
Activated Macrophages ◽  
Virus Expression ◽  
Murine Leukemia

scholarly journals Expression of human adenosine deaminase from various strong promoters after gene transfer into human hematopoietic cell lines

Blood ◽  
1989 ◽  
Vol 74 (2) ◽  
pp. 876-881 ◽  
Author(s):  
RA Hock ◽  
AD Miller ◽  
WR Osborne
Keyword(s):  
Gene Expression ◽  
Cell Line ◽  
Adenosine Deaminase ◽  
Cell Lines ◽  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
K562 Cells ◽  
Moloney Murine Leukemia Virus ◽  
Hematopoietic Cell Lines ◽  
Murine Leukemia

Adenosine deaminase (ADA) deficiency is associated with a fatal severe combined immunodeficiency. Because most patients do not have a suitable marrow donor, the introduction of a normal ADA gene into the patient's marrow cells is a potentially useful alternative therapy. To identify vectors that provide optimal gene expression in human hematopoietic cells, we investigated retroviral vectors containing the ADA gene under the transcriptional control of the promoter/enhancers of Moloney murine leukemia virus, the simian virus 40 early region, the cytomegalovirus immediate-early gene, the lymphotropic papovavirus, and the human beta- globin gene. ADA expression from these vectors was monitored in the ADA- human histiocytic lymphoma cell line DHL-9, and in the multipotential chronic myeloid leukemia cell line K562. ADA expression in infected K562 cells was also measured after induction of megakaryoblastic differentiation by phorbol ester, and after induction of erythroid differentiation by sodium n-butyrate or hemin. In these hematopoietic cell lines, the vectors that contained ADA controlled by either the Moloney murine leukemia virus promoter (LASN) or the cytomegalovirus promoter (LNCA) expressed ADA at much higher levels than the other vectors tested. Furthermore, in K562 cells infected with LASN and LNCA vectors, induction of terminal differentiation resulted in the same or higher level expression of ADA. These cell lines have permitted the evaluation of transduced gene expression in proliferating and differentiating hematopoietic cells that provide a model for bone marrow-targeted gene therapy.

scholarly journals The U3 Region of Moloney Murine Leukemia Virus Contains Position-independent Cis-acting Sequences Involved in the Nuclear Export of Full-length Viral Transcripts

2014 ◽  
Vol 289 (29) ◽  
pp. 20158-20169 ◽  
Author(s):  
Natalia A. Volkova ◽  
Elena G. Fomina ◽  
Viktoryia V. Smolnikova ◽  
Natalia A. Zinovieva ◽  
Igor K. Fomin
Keyword(s):  
Murine Leukemia Virus ◽  
Nuclear Export ◽  
Leukemia Virus ◽  
Full Length ◽  
Moloney Murine Leukemia Virus ◽  
Cis Acting ◽  
Murine Leukemia
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