Synthesis of photolabile transcription initiators and preparation of photocleavable functional RNA by transcription.
Two new photolabile adenosine-containing transcription initiators with terminal thiol and amino functionalities are chemically synthesized. Transcription in the presence of the transcription initiators under the T7 phi2.5 promoter produces 5' thiol- and amino-functionalized RNA conjugated by a photocleavable (PC) linker. Further RNA functionalization with biotin may be achieved through acyl transfer reactions from either biotinyl AMP to the RNA thiol group or biotin NHS to the RNA amino group. Photocleavage of the PC linker displays relatively fast kinetics with a half-life of 4-5 min. The availability of these transcription initiators makes new photolabile RNA accessible for affinity purification of RNA, in vitro selection of functional RNAs, and functional RNA caging.
Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, MS 39406-5043, USA. Faqing.Huang@usm.edu
Faqing
F
Huang
Yongliang
Y
Shi
D000241:Adenosine / Q000737:chemistry; D000249:Adenosine Monophosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D001710:Biotin / Q000031:analogs & derivatives / Q000737:chemistry; D006207:Half-Life; D007700:Kinetics; D010782:Photolysis; D012313:RNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013388:Succinimides / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014158:Transcription, Genetic / Q000187:drug effects*; D014466:Ultraviolet Rays
D013388:Succinimides; D013438:Sulfhydryl Compounds; C033788:biotinyl N-hydroxysuccinimide ester; D000249:Adenosine Monophosphate; D012313:RNA; D001710:Biotin; D000241:Adenosine