title: Generation of CCR5-defective CD34 cells from ZFN-driven stop codon-integrated mesenchymal stem cell clones.
abstract: BACKGROUND
  Homozygous 32-bp deletion of the chemokine receptor 5 gene (CCR5) is associated with resistance to human immunodeficiency virus (HIV) infection, while heterozygosity delays HIV progression. Bone marrow transplantation (BMT) from a 32/32 donor has been shown to cure an HIV-infected patient. However, the rarity of this mutation and the safety risks associated with current BMT protocols are the major obstacles to this treatment. Zinc finger nuclease (ZFN) targeting is a powerful method for achieving genomic disruption at specific DNA sites of interest.
  
  
  RESULTS
  Taking advantage of the self-renewal and plasticity properties of stem cells, in this study, we successfully generated isogenic and six-cell clones of bone marrow-derived mesenchymal stem cells that carry the stop codon of the CCR5 gene by using a ZFN-mediated homology-directed repair technique. These cells were expandable for more than 5 passages, and thus show potential to serve as an individual's cell factory. When Oct4 was overexpressed, the mutated cells robustly converted to CD34+ progenitor cells.
  
  
  CONCLUSIONS
  We here reported the novel approach on generation of patients own CD34 cells from high fidelity ZFN-mediated HDR MSC clones. We believe that our approach will be beneficial in future HIV treatment.
authors.affiliation: Department of Medicine, Molecular and Cell Biology Unit, Lerdsin General Hospital, Bangkok, Thailand. kmanotham@hotmail.com.
authors.forename: Krissanapong
authors.initials: K
authors.lastname: Manotham
authors.affiliation: Department of Medicine, Molecular and Cell Biology Unit, Lerdsin General Hospital, Bangkok, Thailand. s_chattong@yahoo.com.
authors.forename: Supreecha
authors.initials: S
authors.lastname: Chattong
authors.affiliation: Department of Medicine, Molecular and Cell Biology Unit, Lerdsin General Hospital, Bangkok, Thailand. sornin@hotmail.com.
authors.forename: Anant
authors.initials: A
authors.lastname: Setpakdee
mesh_terms: D018388:Codon, Terminator / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D019713:Receptors, CCR5 / Q000235:genetics* / Q000378:metabolism; D059767:Recombinational DNA Repair; D016335:Zinc Fingers
chemical_list: C584933:CCR5 protein, human; D018388:Codon, Terminator; D019713:Receptors, CCR5