Synthesis of symmetrical thiol-adenosine conjugate and 5' thiol-RNA preparation by efficient one-step transcription
1Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, MS 39406-5043, USA. Faqing.Huang@usm.edu
Abstract:
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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