title: Lentiviral vectors transcriptionally targeted to hematopoietic cells by WASP gene proximal promoter sequences.
abstract: The development of vectors that express a therapeutic transgene efficiently and specifically in hematopoietic cells (HCs) is an important goal for gene therapy of hematological disorders. In order to achieve this, we used a 500 bp fragment from the proximal WASP gene promoter to drive the expression of the WASP cDNA in the context of a self-inactivating lentiviral vector. Single-round transduction of WASp-deficient herpesvirus saimiri (HVS)-immortalized cells as well as primary allospecific T cells from Wiskott-Aldrich syndrome (WAS) patients with this vector (WW) resulted in expression levels similar to those of control cells. Non-HCs were transduced with similar efficiency, but the levels of WASp were 135-350 times lower than those achieved in HCs. Additionally, transduction of WASp-deficient cells with WW conferred a selective growth advantage in vitro. Therefore, lentiviral vectors incorporating proximal promoter sequences from the WASP gene confer hematopoietic-specific, and physiological protein expression.
authors.affiliation: IPB 'López Neyra' CSIC, Granada, Spain.
authors.forename: F
authors.initials: F
authors.lastname: Martín
authors.forename: M Ga
authors.initials: MG
authors.lastname: Toscano
authors.forename: M
authors.initials: M
authors.lastname: Blundell
authors.forename: C
authors.initials: C
authors.lastname: Frecha
authors.forename: G K
authors.initials: GK
authors.lastname: Srivastava
authors.forename: M
authors.initials: M
authors.lastname: Santamaría
authors.forename: A J
authors.initials: AJ
authors.lastname: Thrasher
authors.forename: I J
authors.initials: IJ
authors.lastname: Molina
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002461:Cell Line, Transformed; D049109:Cell Proliferation; D003114:Colony-Forming Units Assay; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D006567:Herpesvirus 2, Saimiriine / Q000235:genetics; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D011401:Promoter Regions, Genetic; D011506:Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism*; D014161:Transduction, Genetic / Q000379:methods; D019076:Transgenes; D014923:Wiskott-Aldrich Syndrome / Q000276:immunology / Q000628:therapy*; D051316:Wiskott-Aldrich Syndrome Protein
chemical_list: D011506:Proteins; C495767:WAS protein, human; C495768:Was protein, mouse; D051316:Wiskott-Aldrich Syndrome Protein