title: [Retroviral transfer of a human beta-globin gene linked to beta locus control region hypersensitive site 2 into murine hematopoietic stem cells].
abstract: OBJECTIVE
  To examine the in vivo properties of retroviral recombinants carrying partially deleted human beta-globin gene (delta beta) and truncated erythroid enhancer (292 bp and 341 bp of 5'HS2) at the mRNA levels following short- and long-term reconstitution in mice with infected marrow cells.
  
  
  METHODS
  First ecotropic virus producer cell lines with higher virus titers were isolated using "ping-pong" procedures. Then the human beta-globin gene was transferred into murine hematopoietic progenitor cells and the integration and expression of transferred gene were analyzed by southern blot and RNase protection assay or RT-PCR.
  
  
  RESULTS
  The virus titers of both recombinants increased obviously after "ping-pong" procedures. The transferred human beta-globin gene was detected in murine CFU-S12 and the expression level was about 0.5%-5% of endogenous mouse alpha-globin gene. In 3 of 14 mice surviving long-term transplanted with bone marrow cells transduced with high-titer virus, bone marrow, spleen and thymus from two mice and bone marrow and spleen from another mouse contained the intact proviral genome. Long-term expression of the transferred gene was seen in one mouse at level of 7% of endogenous murine alpha-globin gene.
  
  
  CONCLUSIONS
  The transferred human beta-globin gene can stably integrate into murine hematopoietic stem cells mediated by retroviral vectors and express in an erythroid-specific manner.
authors.affiliation: National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing 100005.
authors.forename: J
authors.initials: J
authors.lastname: Zhai
authors.forename: D
authors.initials: D
authors.lastname: Liu
authors.forename: J
authors.initials: J
authors.lastname: Wang
mesh_terms: D016475:3T3 Cells / Q000378:metabolism; D000818:Animals; D005260:Female; D018014:Gene Transfer Techniques; D005914:Globins / Q000096:biosynthesis* / Q000235:genetics*; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D012190:Retroviridae / Q000235:genetics; D016662:Virus Integration
chemical_list: D005914:Globins