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@ARTICLE{AlJanahi:169364,
      author       = {A. A. AlJanahi and C. R. Lazzarotto and S. Chen and T.-H.
                      Shin and S. Cordes and X. Fan and I. Jabara and Y. Zhou and
                      D. J. Young and B.-C. Lee and K.-R. Yu and Y. Li and B. Toms
                      and I. Tunc and S. G. Hong and L. L. Truitt and J. Klermund
                      and G. Andrieux$^*$ and M. Y. Kim and T. Cathomen and S.
                      Gill and S. Q. Tsai and C. E. Dunbar},
      title        = {{P}rediction and {V}alidation of {H}ematopoietic {S}tem and
                      {P}rogenitor {C}ell {O}ff-{T}arget {E}diting in
                      {T}ransplanted {R}hesus {M}acaques.},
      journal      = {Molecular therapy},
      volume       = {30},
      number       = {1},
      issn         = {1525-0016},
      address      = {New York, NY},
      publisher    = {Nature Publ. Group},
      reportid     = {DKFZ-2021-01444},
      pages        = {209-222},
      year         = {2022},
      note         = {2022 Jan 5;30(1):209-222},
      abstract     = {The programmable nuclease technology CRISPR/Cas9 has
                      revolutionized gene editing in the last decade. Due to the
                      risk of off-target editing, accurate and sensitive methods
                      for off-target characterization are crucial prior to
                      applying CRISPR/Cas9 therapeutically. Here, we utilized a
                      rhesus macaque model to compare the predictive values of
                      CIRCLE-seq, an in vitro off-target prediction method, with
                      in silico prediction (ISP) based solely on genomic sequence
                      comparisons. We use AmpliSeq HD error-corrected sequencing
                      to validate off-target sites predicted by CIRCLE-seq and ISP
                      for a CD33 gRNA with thousands of off-target sites predicted
                      by ISP and CIRCLE-seq. We found poor correlation between the
                      sites predicted by the two methods. When almost 500 sites
                      predicted by each method were analyzed by error-corrected
                      sequencing of hematopoietic cells following transplantation,
                      19 off-target sites revealed insertion/deletion mutations.
                      Of these sites, 8 were predicted by both methods, 8 by
                      CIRCLE-seq only, and 3 by ISP only. The levels of cells with
                      these off-target edits exhibited no expansion or abnormal
                      behavior in vivo in animals followed for up to 2 years. In
                      addition, we utilized an unbiased method termed CAST-Seq to
                      search for translocations between the on-target site and
                      off-target sites present in animals following
                      transplantation, detecting one specific translocation that
                      persisted in blood cells for at least one year following
                      transplantation. In conclusion, neither CIRCLE-seq or ISP
                      predicted all sites, and a combination of careful gRNA
                      design, followed by screening for predicted off-target sites
                      in target cells by multiple methods may be required for
                      optimizing safety of clinical development.},
      cin          = {FR01},
      ddc          = {610},
      cid          = {I:(DE-He78)FR01-20160331},
      pnm          = {899 - ohne Topic (POF4-899)},
      pid          = {G:(DE-HGF)POF4-899},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:34174439},
      doi          = {10.1016/j.ymthe.2021.06.016},
      url          = {https://inrepo02.dkfz.de/record/169364},
}