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CRISPR-Mediated Editing of the B Cell Receptor in Primary Human B Cells.
Greiner, Vera; Bou Puerto, Regina; Liu, Suying; Herbel, Christoph; Carmona, Ellese M; Goldberg, Michael S.
Afiliación
  • Greiner V; Department of Cancer Immunology & Virology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Microbiology & Immunobiology, Harvard Medical School, Boston, MA 02215, USA.
  • Bou Puerto R; Department of Cancer Immunology & Virology, Dana-Farber Cancer Institute, Boston, MA 02215, USA.
  • Liu S; Department of Cancer Immunology & Virology, Dana-Farber Cancer Institute, Boston, MA 02215, USA.
  • Herbel C; Department of Hematology-Oncology and Cancer Biology, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA.
  • Carmona EM; Department of Cancer Immunology & Virology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Microbiology & Immunobiology, Harvard Medical School, Boston, MA 02215, USA.
  • Goldberg MS; Department of Cancer Immunology & Virology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Microbiology & Immunobiology, Harvard Medical School, Boston, MA 02215, USA. Electronic address: michael_goldberg1@dfci.harvard.edu.
iScience ; 12: 369-378, 2019 Feb 22.
Article en En | MEDLINE | ID: mdl-30769282
Vaccination approaches have generally focused on the antigen rather than the resultant antibodies generated, which differ greatly in quality and function between individuals. The ability to replace the variable regions of the native B cell receptor (BCR) heavy and light chain loci with defined recombined sequences of a preferred monoclonal antibody could enable curative adoptive cell transfer. We report CRISPR-mediated homologous recombination (HR) into the BCR of primary human B cells. Ribonucleoprotein delivery enabled editing at the model CXCR4 locus, as demonstrated by T7E1 assay, flow cytometry, and TIDE analysis. Insertion via HR was confirmed by sequencing, cross-boundary PCR, and restriction digest. Optimized conditions were used to achieve HR at the BCR variable heavy and light chains. Insertion was confirmed at the DNA level, and transgene expression from the native BCR promoters was observed. Reprogramming the specificity of antibodies in the genomes of B cells could have clinical importance.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Tipo de estudio: Prognostic_studies Idioma: En Revista: IScience Año: 2019 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Tipo de estudio: Prognostic_studies Idioma: En Revista: IScience Año: 2019 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos