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A solid-phase transfection platform for arrayed CRISPR screens.
Serçin, Özdemirhan; Reither, Sabine; Roidos, Paris; Ballin, Nadja; Palikyras, Spyridon; Baginska, Anna; Rein, Katrin; Llamazares, Maria; Halavatyi, Aliaksandr; Winter, Hauke; Muley, Thomas; Jurkowska, Renata Z; Abdollahi, Amir; Zenke, Frank T; Neumann, Beate; Mardin, Balca R.
Afiliação
  • Serçin Ö; BioMed X Innovation Center, Heidelberg, Germany.
  • Reither S; Advanced Light Microscopy Facility, European Molecular Biology Laboratory, Heidelberg, Germany.
  • Roidos P; BioMed X Innovation Center, Heidelberg, Germany.
  • Ballin N; BioMed X Innovation Center, Heidelberg, Germany.
  • Palikyras S; BioMed X Innovation Center, Heidelberg, Germany.
  • Baginska A; BioMed X Innovation Center, Heidelberg, Germany.
  • Rein K; BioMed X Innovation Center, Heidelberg, Germany.
  • Llamazares M; BioMed X Innovation Center, Heidelberg, Germany.
  • Halavatyi A; Advanced Light Microscopy Facility, European Molecular Biology Laboratory, Heidelberg, Germany.
  • Winter H; Department of Surgery, Thoraxklinik at University Hospital Heidelberg, Heidelberg, Germany.
  • Muley T; Translational Lung Research Center (TLRC) Heidelberg, Member of the German Center for Lung Research (DZL), Heidelberg, Germany.
  • Jurkowska RZ; Translational Lung Research Center (TLRC) Heidelberg, Member of the German Center for Lung Research (DZL), Heidelberg, Germany.
  • Abdollahi A; Thoraxklinik at University Hospital Heidelberg, Heidelberg, Germany.
  • Zenke FT; BioMed X Innovation Center, Heidelberg, Germany.
  • Neumann B; Division of Molecular and Translational Radiation Oncology, National Center for Tumor Diseases (NCT), and German Cancer Research Center (DKFZ), Heidelberg University Hospital, Heidelberg, Germany.
  • Mardin BR; Clinical Cooperation Unit Translational Radiation Oncology, German Cancer Consortium (DKTK) Core Center Heidelberg, Heidelberg, Germany.
Mol Syst Biol ; 15(12): e8983, 2019 12.
Article em En | MEDLINE | ID: mdl-31885201
ABSTRACT
Arrayed CRISPR-based screens emerge as a powerful alternative to pooled screens making it possible to investigate a wide range of cellular phenotypes that are typically not amenable to pooled screens. Here, we describe a solid-phase transfection platform that enables CRISPR-based genetic screens in arrayed format with flexible readouts. We demonstrate efficient gene knockout upon delivery of guide RNAs and Cas9/guide RNA ribonucleoprotein complexes into untransformed and cancer cell lines. In addition, we provide evidence that our platform can be easily adapted to high-throughput screens and we use this approach to study oncogene addiction in tumor cells. Finally demonstrating that the human primary cells can also be edited using this method, we pave the way for rapid testing of potential targeted therapies.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA Guia de Cinetoplastídeos / Edição de Genes / Neoplasias Limite: Humans Idioma: En Revista: Mol Syst Biol Assunto da revista: BIOLOGIA MOLECULAR / BIOTECNOLOGIA Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Alemanha

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA Guia de Cinetoplastídeos / Edição de Genes / Neoplasias Limite: Humans Idioma: En Revista: Mol Syst Biol Assunto da revista: BIOLOGIA MOLECULAR / BIOTECNOLOGIA Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Alemanha