Variable stability of a selectable provirus after retroviral vector gene transfer into human cells.
AUTOR(ES)
Jolly, D J
RESUMO
Human lymphoblasts deficient in the enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) were infected with an amphotropic helper-free retroviral vector expressing human HPRT cDNA. The stability and expression of the HPRT provirus in five cell lines with different proviral integration sites were examined by determining HPRT mutation and reversion frequencies and by blot hybridization studies. Mutation to the HPRT-negative phenotype occurred at frequencies of approximately 4 X 10(-5) to 3 X 10(-6) per generation. Most mutations in each of the five cell lines were associated with partial or complete deletions or rearrangements of the provirus. Several mutants retained a grossly intact HPRT provirus, and in one such mutant HPRT shutdown resulted from a revertible epigenetic mechanism that was not associated with global changes in proviral methylation. Therefore, mutation and shutdown of the HPRT provirus in human lymphoblasts result from mechanisms similar to those reported for several other avian and mammalian replication-competent retroviruses.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=367625Documentos Relacionados
- An improved retroviral vector for gene transfer into undifferentiated cells.
- Production of human glucocerebrosidase in mice after retroviral gene transfer into multipotential hematopoietic progenitor cells.
- Expression of the human beta-globin gene after retroviral transfer into murine erythroleukemia cells and human BFU-E cells.
- Retroviral mediated gene transfer into bone marrow progenitor cells: use of beta-galactosidase as a selectable marker.
- Transient expression of the chicken lysozyme gene after transfer into human cells.