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@ARTICLE{Nasri:153073,
      author       = {M. Nasri and P. Mir$^*$ and B. Dannenmann and D. Amend and
                      T. Skroblyn and Y. Xu and K. Schulze-Osthoff$^*$ and M.
                      Klimiankou and K. Welte and J. Skokowa},
      title        = {{F}luorescent labeling of {CRISPR}/{C}as9 {RNP} for gene
                      knockout in {HSPC}s and i{PSC}s reveals an essential role
                      for {GADD}45b in stress response.},
      journal      = {Blood advances},
      volume       = {3},
      number       = {1},
      issn         = {2473-9537},
      address      = {Washington, DC},
      publisher    = {American Society of Hematology},
      reportid     = {DKFZ-2020-00155},
      pages        = {63 - 71},
      year         = {2019},
      abstract     = {CRISPR/Cas9-mediated gene editing of stem cells and primary
                      cell types has several limitations for clinical
                      applications. The direct delivery of ribonucleoprotein (RNP)
                      complexes consisting of Cas9 nuclease and guide RNA (gRNA)
                      has improved DNA- and virus-free gene modifications, but it
                      does not enable the essential enrichment of the gene-edited
                      cells. Here, we established a protocol for the fluorescent
                      labeling and delivery of CRISPR/Cas9-gRNA RNP in primary
                      human hematopoietic stem and progenitor cells (HSPCs) and
                      induced pluripotent stem cells (iPSCs). As a proof of
                      principle for genes with low-abundance transcripts and
                      context-dependent inducible expression, we successfully
                      deleted growth arrest and DNA-damage-inducible β (GADD45B).
                      We found that GADD45B is indispensable for DNA damage
                      protection and survival in stem cells. Thus, we describe an
                      easy and efficient protocol of DNA-free gene editing of
                      hard-to-target transcripts and enrichment of gene-modified
                      cells that are generally difficult to transfect.},
      keywords     = {Antigens, Differentiation (NLM Chemicals) / GADD45B
                      protein, human (NLM Chemicals) / Macromolecular Substances
                      (NLM Chemicals) / RNA, Guide (NLM Chemicals) /
                      Ribonucleoproteins (NLM Chemicals)},
      cin          = {L801},
      ddc          = {610},
      cid          = {I:(DE-He78)L801-20160331},
      pnm          = {899 - ohne Topic (POF3-899)},
      pid          = {G:(DE-HGF)POF3-899},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:30622144},
      pmc          = {pmc:PMC6325296},
      doi          = {10.1182/bloodadvances.2017015511},
      url          = {https://inrepo02.dkfz.de/record/153073},
}