title: TALEN-mediated genetic tailoring as a tool to analyze the function of acquired mutations in multiple myeloma cells.
abstract: Multiple myeloma (MM) is a clonal plasma cell malignancy that is initiated by a number of mutations and the process of disease progression is characterized by further acquisition of mutations. The identification and functional characterization of these myelomagenic mutations is necessary to better understand the underlying pathogenic mechanisms in this disease. Recent advancements in next-generation sequencing have made the identification of most of these mutations a reality. However, the functional characterization of these mutations has been hampered by the lack of proper and efficient tools to dissect these mutations. Here we explored the possible utility of transcription activator-like effector nuclease (TALEN) genome engineering technology to tailoring the genome of MM cells. To test this possibility, we targeted the HPRT1 gene and found that TALENs are a very robust and efficient genome-editing tool in MM cells. Using cotransfected green fluorescent protein as an enrichment marker, single-cell subclones with desirable TALEN modifications in the HPRT1 gene were obtained in as little as 3-4 weeks of time. We believe that TALENs will greatly facilitate the functional study of somatic mutations in MM as well as other cancers.
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Rochester, MN, USA.
authors.forename: X
authors.initials: X
authors.lastname: Wu
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, MN, USA.
authors.forename: P R
authors.initials: PR
authors.lastname: Blackburn
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Rochester, MN, USA.
authors.forename: R C
authors.initials: RC
authors.lastname: Tschumper
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, MN, USA.
authors.forename: S C
authors.initials: SC
authors.lastname: Ekker
authors.affiliation: 1] Department of Immunology, Mayo Clinic College of Medicine, Rochester, MN, USA [2] Department of Internal Medicine, Mayo Clinic College of Medicine, Rochester, MN, USA.
authors.forename: D F
authors.initials: DF
authors.lastname: Jelinek
mesh_terms: D001483:Base Sequence; D003851:Deoxyribonucleases / Q000235:genetics* / Q000378:metabolism; D005260:Female; D055786:Gene Knockout Techniques / Q000379:methods; D018390:Gene Targeting; D005818:Genetic Engineering; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000201:enzymology* / Q000235:genetics*; D009154:Mutation; D014407:Tumor Cells, Cultured
chemical_list: D007041:Hypoxanthine Phosphoribosyltransferase; D003851:Deoxyribonucleases