Optogenetic Control of Nuclear Protein Import in Living Cells Using Light-Inducible Nuclear Localization Signals (LINuS).
Many biological processes are regulated by the timely import of specific proteins into the nucleus. The ability to spatiotemporally control the nuclear import of proteins of interest therefore allows study of their role in a given biological process as well as controlling this process in space and time. The light-inducible nuclear localization signal (LINuS) was developed based on a natural plant photoreceptor that reversibly triggers the import of proteins of interest into the nucleus with blue light. Each LINuS is a small, genetically encoded domain that is fused to the protein of interest at the N or C terminus. These protocols describe how to carry out initial microscopy-based screening to assess which LINuS variant works best with a protein of interest. © 2016 by John Wiley & Sons, Inc.
Center for Quantitative Analysis of Molecular and Cellular Biosystems (BioQuant), University of Heidelberg, Heidelberg, Germany.
Pierre
P
Wehler
Center for Quantitative Analysis of Molecular and Cellular Biosystems (BioQuant), University of Heidelberg, Heidelberg, Germany.|Department of Bioinformatics and Functional Genomics, Institute for Pharmacy and Biotechnology.|Department of Theoretical Bioinformatics, German Cancer Research Center, Heidelberg, Germany.
Dominik
D
Niopek
Center for Quantitative Analysis of Molecular and Cellular Biosystems (BioQuant), University of Heidelberg, Heidelberg, Germany.|Department of Bioinformatics and Functional Genomics, Institute for Pharmacy and Biotechnology.|Department of Theoretical Bioinformatics, German Cancer Research Center, Heidelberg, Germany.
Roland
R
Eils
Center for Quantitative Analysis of Molecular and Cellular Biosystems (BioQuant), University of Heidelberg, Heidelberg, Germany.
Barbara
B
Di Ventura
D021581:Active Transport, Cell Nucleus / Q000528:radiation effects; D000818:Animals; D008027:Light; D019913:Nuclear Localization Signals / Q000737:chemistry* / Q000528:radiation effects*; D009687:Nuclear Proteins / Q000378:metabolism; D062308:Optogenetics / Q000379:methods*; D015202:Protein Engineering / Q000379:methods*
D019913:Nuclear Localization Signals; D009687:Nuclear Proteins
LOV2; nuclear import; optogenetics; protein engineering; synthetic biology