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Transcript
Light‐Based Technologies Light Activation • Induce Transcription • Transcription factors • Induce Translation • Activate Proteins • Protein dimerization • Protein translocation Light-Controlled Circuits Phytochromes – The PhyB-Pif6 System • Native to plants • Used to control protein translocation • Advantages • Red Light • Photoswitchable • Long half‐life (hours) A Levskaya et al. Nature 461, 997-1001 (2009) • Disadvantages • Requires bilin co‐factor • Short half‐life (seconds) Phytochromes – The PhyB-Pif6 System Beyer HM et al. ACS Synth Biol 2015 Ahead of Print Phytochromes – The PhyB-Pif6 System Konrad Müller et al. Nucl. Acids Res. 2013;41:e77 Used in: Bacteria, Yeast, Cell Culture Cryptochromes – The Cry2-CIB1 System • Native to plants and animals • Used to control protein interactions • Advantages • Blue Light • FAD co‐factor • Disadvantages • Short half‐life (minutes) Kennedy et al. Nature Methods 7, 973-975 (2010) Cryptochromes – The Cry2-CIB1 System Lui et al. PLoS One 7 (11): e50738 (2012) Cryptochromes – The Cry2-CIB1 System Polstein et al. Nature Chemical Biology 11, 198-200 (2015) LOV Domains – The LOV2 System • Domain found in phototropin of oat • Used to control protein activity • Advantages • Blue Light • Flavin cofactor • Disadvantages • Short half‐life (seconds) Yi et al. Nature 461, 104-108 (2009) LOV Domains – The FKF1-GIGANTEA System • Found in mouse‐cress • Used to control transcription • Advantages • Blue Light • Flavin cofactor • Long half‐life (days) Polstein et al. JACS 134, 16480-16483 (2012) LOV Domains – The EL222 System • Found in bacteria • Used to control transcription • Advantages • Blue Light • Flavin cofactor • Unique binding site • Disadvantages • Short half‐life (seconds) Motta-Mena et al. Nature Chemical Biology 10, 196-202 (2014) LOV Domains – The EL222 System Motta-Mena et al. Nature Chemical Biology 10, 196-202 (2014) LOV Domains – The VIVID System • Found in fungus • Used to control transcription • Advantages • Blue Light • FAD Cofactor • Long half‐life (hours) • Disadvantages • Requires strong activator Wang et al. Nature Methods 9, 266-269 (2012) UV Photoreceptors – The UVR8-COP1 System • Found in arabidopsis • Used to control protein interactions • Disadvantages • UV light Crefcoeur et al. Nature Communications 4, 1779 (2013) Light Activation System Species Used For Half-Life Kinetics Phytochromes Bacteria, Cell culture Protein Localization and Transcription Sub-second Cryptochromes Yeast, Cell culture, Zebrafish Protein Localization, Protein Assembly, and Transcription Minutes LOV2 Cell culture Protein Localization, Seconds and Protein Activation FKF1 + GIGANTEA Cell culture Protein Localization and Transcription Days VIVID Cell culture, Mice Transcription Hours EL222 Cell culture, Zebrafish Transcription Sub-second UVR8 + COP1 Cell culture Protein Localization, Transcription Hours
