Murine retroviral but not human cellular promoters induce in vivo erythroid-specific deregulation that can be partially prevented by insulators.

摘要: We are developing lentiviral vectors for gene therapy of red blood cell disorders that co-express a transgene in an erythroid-specific manner and the O6-methylguanine-DNA-methyltransferase (MGMT) selective constitutive way. report transduction murine hematopoietic stem cells (HSCs) with human phosphoglycerate kinase promoter-based vector at low multiplicity infection (MOI) does not result vivo expansion presence alkylating agents. In contrast, by replacing this cellular promoter powerful retroviral-derived myeloproliferative sarcoma virus enhancer, negative control region-deleted, dl587rev primer-binding site substituted promoter, allowed efficient chemoprotection transduced HSCs MOI. However, interacted erythroid HS40/ankyrin enhancer/promoter driving green fluorescent protein, leading to unexpected loss specificity. A partial restoration tissue-specific expression was obtained interposition insulator sequences between units. Alternatively, we found strong elongation factor1-α allows similar but without any deregulation absence insulators. These data demonstrate level induced is correlated its transcriptional activity.