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

An amphotropic retrovirus packaging cell line was constructed in which the gag, pol, and env genes of the helper virus are separated on two different plasmids and in which the packaging signals and 3' long terminal repeats are removed. To do this, a plasmid containing the Moloney murine leukemia virus gag and pol gene was transfected into NIH 3T3 cells, and a plasmid containing the 4070A amphotropic env gene was transfected into one of the resulting clones which produced a high level of reverse transcriptase. A clone producing a high level of amphotropic env protein (GP + envAm12) was then isolated. When transfected into GP + envAm12 cells, titers of the retroviral vector N2, containing a neomycin resistance gene, ranged from 10(2) to greater than 10(6) CFU/ml on 3T3 cells, from 1.3 x 10(4) to 2.7 x 10(5) CFU/ml on HeLa cells, and from 1.0 x 10(2) to 6.0 x 10(3) CFU/ml on K562 cells when assayed by G418 resistance. These titers were comparable to titers obtained using the PA317 cell line. Tests for the safety of the GP + envAm12 packaging line showed no evidence for the generation of wild-type virus. Thus, the efficiency and safety of the GP + envAm12 cell line in gene transfer into human cells may provide an optimal system for experiments whose goal is human gene therapy.
 ...
PMID:Construction and use of a safe and efficient amphotropic packaging cell line. 246 7

A retrovirus packaging cell line was constructed by using portions of the Moloney murine leukemia virus in which the gag, pol, and env genes of the helper virus were separated onto two different plasmids and in which the psi packaging signal and 3' long terminal repeat were removed. The plasmid containing the gag and pol genes and the plasmid containing the env gene were cotransfected into NIH 3T3 cells. Clones that produced high levels of reverse transcriptase and env protein were tested for their ability to package the replication-defective retrovirus vectors delta neo and N2. One of the gag-pol and env clones (GP+E-86) was able to transfer G418 resistance to recipient cells at a titer of as high as 1.7 X 10(5) when it was used to package delta neo and as high as 4 X 10(6) when it was used to package N2. Supernatants of clones transfected with the intact parent gag-pol-env plasmid 3P0 had comparable titers (as high as 6.5 X 10(4) with delta neo; as high as 1.7 X 10(5) with N2). Tests for recombination events that might result in intact retrovirus showed no evidence for the generation of replication-competent virus. These results suggest that gag, pol, and env, when present on different plasmids, may provide an efficient and safe packaging line for use in retroviral gene transfer.
 ...
PMID:A safe packaging line for gene transfer: separating viral genes on two different plasmids. 283 75