Sequence-independent and reversible photocontrol of transcription/expression systems using a photosensitive nucleic acid binder.
To understand non-trivial biological functions, it is crucial to develop minimal synthetic models that capture their basic features. Here, we demonstrate a sequence-independent, reversible control of transcription and gene expression using a photosensitive nucleic acid binder (pNAB). By introducing a pNAB whose affinity for nucleic acids is tuned by light, in vitro RNA production, EGFP translation, and GFP expression (a set of reactions including both transcription and translation) were successfully inhibited in the dark and recovered after a short illumination at 365 nm. Our results indicate that the accessibility of the protein machinery to one or several nucleic acid binding sites can be efficiently regulated by changing the conformational/condensation state of the nucleic acid (DNA conformation or mRNA aggregation), thus regulating gene activity in an efficient, reversible, and sequence-independent manner. The possibility offered by our approach to use light to trigger various gene expression systems in a system-independent way opens interesting perspectives to study gene expression dynamics as well as to develop photocontrolled biotechnological procedures.
Estévez-Torres
André
A
Department of Physics, Kyoto University, Kyoto 606-8502, Japan.
Crozatier
Cécile
C
Diguet
Antoine
A
Hara
Tomoaki
T
Saito
Hirohide
H
Yoshikawa
Kenichi
K
Baigl
Damien
D
D017122:Bacteriophage T4 / Q000235:genetics; D001583:Benzoxazoles / Q000737:chemistry; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics*; D005786:Gene Expression Regulation / Q000528:radiation effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D011806:Quinolinium Compounds / Q000737:chemistry; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D014158:Transcription, Genetic; D014466:Ultraviolet Rays
D001583:Benzoxazoles; D004279:DNA, Viral; D000644:Quaternary Ammonium Compounds; D011806:Quinolinium Compounds; D012333:RNA, Messenger; C075296:1,1'-((4,4,7,7-tetramethyl)-4,7-diazaundecamethylene)bis-4-(3-methyl-2,3-dihydro(benzo-1,3-oxazole)-2-methylidene)quinolinium; D049452:Green Fluorescent Proteins; C006664:4-azobenzenetrimethylammonium