Genomic rearrangements of retroviral vectors carrying two genes in F9 EC cells.
We have used two classes of double-expression retroviral vectors for the expression of foreign genetic information in embryonal carcinoma cell lines. The splice-vector pM5neo takes advantage of mutated sequences that mediate an LTR-driven expression in F9 EC cells. The second vector (pXT1 type) uses an internal HSV-tk promoter as the control element for the transcription of the second gene. Genomic analysis of DNA from infected F9 cell lines revealed that most of the proviruses have rearranged upon integration into the host genome. This reorganization always included the nonselected gene and is sequence independent, but depends on the selective pressure applied. No retroviral genomic rearrangements were observed in F9 cells infected with pM5 proviruses carrying only the neo resistance gene. On the contrary, gross rearrangements were found in cells infected with parental pXT1 retroviruses. In both vectors the transcriptional activity was very low. A direct correlation between selective pressure, proviral reorganization, and transcription was observed.
Breuer
B
B
Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Steuer
B
B
Alonso
A
A
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D015870:Gene Expression; D015321:Gene Rearrangement; D005814:Genes, Viral; D005822:Genetic Vectors; D051379:Mice; D011533:Proviruses / Q000235:genetics; D012313:RNA / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D013724:Teratoma / Q000235:genetics; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured / Q000378:metabolism
D004274:DNA, Recombinant; C039919:RNA, recombinant; D012313:RNA