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Creating New ß-Globin-Expressing Lentiviral Vectors by High-Resolution Mapping of Locus Control Region Enhancer Sequences.
Morgan, Richard A; Ma, Feiyang; Unti, Mildred J; Brown, Devin; Ayoub, Paul George; Tam, Curtis; Lathrop, Lindsay; Aleshe, Bamidele; Kurita, Ryo; Nakamura, Yukio; Senadheera, Shantha; Wong, Ryan L; Hollis, Roger P; Pellegrini, Matteo; Kohn, Donald B.
Afiliación
  • Morgan RA; Charles R. Drew University of Medicine and Science, Los Angeles, CA 90059, USA.
  • Ma F; Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
  • Unti MJ; Molecular Biology Institute Interdepartmental Doctoral Program, University of California, Los Angeles, Los Angeles, CA 90095, USA.
  • Brown D; Department of Microbiology, Immunology & Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
  • Ayoub PG; Department of Microbiology, Immunology & Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
  • Tam C; Department of Microbiology, Immunology & Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
  • Lathrop L; Department of Microbiology, Immunology & Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
  • Aleshe B; Department of Microbiology, Immunology & Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
  • Kurita R; Department of Microbiology, Immunology & Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
  • Nakamura Y; Cell Engineering Division, RIKEN BioResource Center, Tsukuba, Ibaraki, Japan.
  • Senadheera S; Cell Engineering Division, RIKEN BioResource Center, Tsukuba, Ibaraki, Japan.
  • Wong RL; Department of Microbiology, Immunology & Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
  • Hollis RP; Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
  • Pellegrini M; Department of Microbiology, Immunology & Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
  • Kohn DB; Molecular Biology Institute Interdepartmental Doctoral Program, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Mol Ther Methods Clin Dev ; 17: 999-1013, 2020 Jun 12.
Article en En | MEDLINE | ID: mdl-32426415
ABSTRACT
Hematopoietic stem cell gene therapy is a promising approach for treating disorders of the hematopoietic system. Identifying combinations of cis-regulatory elements that do not impede packaging or transduction efficiency when included in lentiviral vectors has proven challenging. In this study, we deploy LV-MPRA (lentiviral vector-based, massively parallel reporter assay), an approach that simultaneously analyzes thousands of synthetic DNA fragments in parallel to identify sequence-intrinsic and lineage-specific enhancer function at near-base-pair resolution. We demonstrate the power of LV-MPRA in elucidating the boundaries of previously unknown intrinsic enhancer sequences of the human ß-globin locus control region. Our approach facilitated the rapid assembly of novel therapeutic ßAS3-globin lentiviral vectors harboring strong lineage-specific recombinant control elements capable of correcting a mouse model of sickle cell disease. LV-MPRA can be used to map any genomic locus for enhancer activity and facilitates the rapid development of therapeutic vectors for treating disorders of the hematopoietic system or other specific tissues and cell types.
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Texto completo: 1 Bases de datos: MEDLINE Idioma: En Revista: Mol Ther Methods Clin Dev Año: 2020 Tipo del documento: Article País de afiliación: Estados Unidos
Texto completo
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Bases de datos: MEDLINE Idioma: En Revista: Mol Ther Methods Clin Dev Año: 2020 Tipo del documento: Article País de afiliación: Estados Unidos