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@ARTICLE{Becker:157427,
      author       = {M. Becker and H. Noll-Puchta and D. Amend and F. Nolte and
                      C. Fuchs and I. Jeremias$^*$ and C. J. Braun$^*$},
      title        = {{CLUE}: a bioinformatic and wet-lab pipeline for
                      multiplexed cloning of custom sg{RNA} libraries.},
      journal      = {Nucleic acids research},
      volume       = {48},
      number       = {13},
      issn         = {0301-5610},
      address      = {Oxford},
      publisher    = {Oxford Univ. Press},
      reportid     = {DKFZ-2020-01622},
      pages        = {e78},
      year         = {2020},
      note         = {#LA:B062#},
      abstract     = {The systematic perturbation of genomes using CRISPR/Cas9
                      deciphers gene function at an unprecedented rate, depth and
                      ease. Commercially available sgRNA libraries typically
                      contain tens of thousands of pre-defined constructs,
                      resulting in a complexity challenging to handle. In
                      contrast, custom sgRNA libraries comprise gene sets of
                      self-defined content and size, facilitating experiments
                      under complex conditions such as in vivo systems. To
                      streamline and upscale cloning of custom libraries, we
                      present CLUE, a bioinformatic and wet-lab pipeline for the
                      multiplexed generation of pooled sgRNA libraries. CLUE
                      starts from lists of genes or pasted sequences provided by
                      the user and designs a single synthetic oligonucleotide pool
                      containing various libraries. At the core of the approach, a
                      barcoding strategy for unique primer binding sites allows
                      amplifying different user-defined libraries from one single
                      oligonucleotide pool. We prove the approach to be
                      straightforward, versatile and specific, yielding uniform
                      sgRNA distributions in all resulting libraries, virtually
                      devoid of cross-contaminations. For in silico library
                      multiplexing and design, we established an easy-to-use
                      online platform at www.crispr-clue.de. All in all, CLUE
                      represents a resource-saving approach to produce numerous
                      high quality custom sgRNA libraries in parallel, which will
                      foster their broad use across molecular biosciences.},
      cin          = {MU01 / B062},
      ddc          = {570},
      cid          = {I:(DE-He78)MU01-20160331 / I:(DE-He78)B062-20160331},
      pnm          = {312 - Functional and structural genomics (POF3-312)},
      pid          = {G:(DE-HGF)POF3-312},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:32479629},
      pmc          = {pmc:PMC7367185},
      doi          = {10.1093/nar/gkaa459},
      url          = {https://inrepo02.dkfz.de/record/157427},
}