title: Restriction of expression of an integrated recombinant retrovirus in primary but not immortalized murine hematopoietic stem cells.
abstract: A recombinant retrovirus (DHFR*-SVADA) in which human adenosine deaminase (ADA) cDNA is transcribed from an internal SV40 promoter was used to infect murine hematopoietic stem and progenitor cells. Human ADA enzyme was not expressed in infected primary murine pluripotent stem cell-derived spleen or progenitor colonies (CFU-GM, CFU-Mix, BFU-E). In contrast, human ADA enzyme activity was readily detected in progenitor colonies derived from immortalized multipotent factor-dependent cells. The level of human enzyme was near endogenous murine enzyme levels and was equivalent in undifferentiated stem cells and differentiated myeloid, erythroid, and mixed colonies. These results indicate that cellular properties other than the stage of differentiation are important in determining the expression of foreign sequences introduced by retroviruses. Cell lines that are immortalized but still capable of induced differentiation may contain factors that abrogate blocks to expression that are manifested in primary hematopoietic stem cells.
authors.affiliation: Division of Hematology-Oncology, Howard Hughes Medical Institute, Boston, MA.
authors.forename: D A
authors.initials: DA
authors.lastname: Williams
authors.forename: B
authors.initials: B
authors.lastname: Lim
authors.forename: E
authors.initials: E
authors.lastname: Spooncer
authors.forename: J
authors.initials: J
authors.lastname: Longtine
authors.forename: T M
authors.initials: TM
authors.lastname: Dexter
mesh_terms: D000243:Adenosine Deaminase / Q000235:genetics; D002454:Cell Differentiation; D004274:DNA, Recombinant; D005786:Gene Expression Regulation; D006133:Growth Substances / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology*; D006801:Humans; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D013539:Simian virus 40 / Q000235:genetics; D014161:Transduction, Genetic
chemical_list: D004274:DNA, Recombinant; D006133:Growth Substances; D012333:RNA, Messenger; D000243:Adenosine Deaminase