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 Phi 2.5 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. (C) 2010 Elsevier Ltd. All rights reserved.