Synthesis of symmetrical thiol-adenosine conjugate and 5′ thiol-RNA preparation by efficient one-step transcription

✍ Scribed by Faqing Huang; Yongliang Shi

• Publisher: Elsevier Science
• Year: 2010
• Tongue: English
• Weight: 445 KB
• Volume: 20
• Category: Article
• ISSN: 0960-894X
• DOI: 10.1016/j.bmcl.2010.08.097

✦ Synopsis

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.