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@ARTICLE{Yudenko:904315,
      author       = {Yudenko, Anna and Smolentseva, Anastasia and Maslov, Ivan
                      and Semenov, Oleg and Goncharov, Ivan M. and Nazarenko, Vera
                      V. and Maliar, Nina L. and Borshchevskiy, Valentin and
                      Gordeliy, Valentin and Remeeva, Alina and Gushchin, Ivan},
      title        = {{R}ational {D}esign of a {S}plit {F}lavin-{B}ased
                      {F}luorescent {R}eporter},
      journal      = {ACS synthetic biology},
      volume       = {10},
      number       = {1},
      issn         = {2161-5063},
      address      = {Washington, DC},
      publisher    = {ACS},
      reportid     = {FZJ-2021-05885},
      pages        = {72 - 83},
      year         = {2021},
      abstract     = {Protein-fragment complementation assays are used
                      ubiquitously for probing protein–protein interactions.
                      Most commonly, the reporter protein is split in two parts,
                      which are then fused to the proteins of interest and can
                      reassemble and provide a readout if the proteins of interest
                      interact with each other. The currently known split
                      fluorescent proteins either can be used only in aerobic
                      conditions and assemble irreversibly, or require addition of
                      exogenous chromophores, which complicates the design of
                      experiments. In recent years, light-oxygen-voltage (LOV)
                      domains of several photoreceptor proteins have been
                      developed into flavin-based fluorescent proteins (FbFPs)
                      that, under some circumstances, can outperform commonly used
                      fluorescent proteins such as GFP. Here, we show that
                      CagFbFP, a small thermostable FbFP based on a LOV
                      domain-containing protein from Chloroflexus aggregans, can
                      serve as a split fluorescent reporter. We use the available
                      genetic and structural information to identify three loops
                      between the conserved secondary structure elements, Aβ-Bβ,
                      Eα-Fα, and Hβ-Iβ, that tolerate insertion of flexible
                      poly-Gly/Ser segments and eventually splitting. We
                      demonstrate that the designed split pairs, when fused to
                      interacting proteins, are fluorescent in vivo in E. coli and
                      human cells and have low background fluorescence. Our
                      results enable probing protein–protein interactions in
                      anaerobic conditions without using exogenous fluorophores
                      and provide a basis for further development of LOV and PAS
                      (Per-Arnt-Sim) domain-based fluorescent reporters and
                      optogenetic tools.},
      cin          = {IBI-7},
      ddc          = {570},
      cid          = {I:(DE-Juel1)IBI-7-20200312},
      pnm          = {5241 - Molecular Information Processing in Cellular Systems
                      (POF4-524)},
      pid          = {G:(DE-HGF)POF4-5241},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {33325704},
      UT           = {WOS:000643603000007},
      doi          = {10.1021/acssynbio.0c00454},
      url          = {https://juser.fz-juelich.de/record/904315},
}