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.

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