Synthesis of symmetrical thiol-adenosine conjugate and 5' thiol-RNA preparation by efficient one-step transcription.
A symmetrical transcription initiator containing two adenosines and conjugated thiol functionality (ThioAMP dimer) is chemically synthesized. Transcription in the presence of ThioAMP dimer under the T7 Φ2.5 promoter yields 5' thiol-labeled RNA (5' HS-RNA) with up to 90% labeling efficiency, depending on the concentration ratio of ThioAMP dimer to ATP. The resulting 5' HS-RNA may be used directly or after thiopropyl Separose 6B affinity column purification. Biotinylation of 5' HS-RNA and formation of gold nanoparticle-RNA nanoplexes are demonstrated.
Huang
Faqing
F
Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, MS 39406-5043, USA. Faqing.Huang@usm.edu
Shi
Yongliang
Y
D000241:Adenosine / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006046:Gold; D007202:Indicators and Reagents; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000737:chemistry; D053758:Nanoparticles; D012313:RNA / Q000138:chemical synthesis / Q000737:chemistry*; D034741:RNA, Small Interfering; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D000077022:Survivin; D014158:Transcription, Genetic / Q000187:drug effects*
C107489:BIRC5 protein, human; D007202:Indicators and Reagents; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D034741:RNA, Small Interfering; D013438:Sulfhydryl Compounds; D000077022:Survivin; D012313:RNA; D001710:Biotin; D006046:Gold; D000241:Adenosine