Unit

UNIT 2.10 Preparation of 5′-Silyl-2′-Orthoester Ribonucleosides for Use in Oligoribonucleotide Synthesis

  1. Stephen A. Scaringe,
  2. David Kitchen,
  3. Robert J. Kaiser,
  4. William S. Marshall

Published Online: 1 MAY 2004

DOI: 10.1002/0471142700.nc0210s16

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Scaringe, S. A., Kitchen, D., Kaiser, R. J. and Marshall, W. S. 2004. Preparation of 5′-Silyl-2′-Orthoester Ribonucleosides for Use in Oligoribonucleotide Synthesis. Current Protocols in Nucleic Acid Chemistry. 16:2.10:2.10.1–2.10.16.

Author Information

  1. Dharmacon Inc., Lafayette, Colorado

Publication History

  1. Published Online: 1 MAY 2004
  2. Published Print: MAR 2004

Abstract

The recent discovery that small interfering RNAs (siRNAs) induce gene suppression in mammalian cells has sparked tremendous interest in using siRNA-based assays and high-throughput screens to study gene function. As a result, research programs at leading academic and commercial institutions have become a substantial and rapidly growing market for synthetic RNA. Important considerations in synthesizing RNA for biological gene function studies are sequence integrity, purity, scalability, and resistance to nucleases; ease of chemical modification, deprotection, and handling; and cost. Of the well-established RNA synthesis methods, 2′-ACE chemistry is the only one that meets all of these criteria. 2′-ACE technology employs a unique class of silyl ethers to protect the 5′-hydroxyl, in combination with an acid-labile orthoester protecting group on the 2′-hydroxyl (2′-ACE). 2′-ACE-protected phosphoramidite monomers are joined using standard solid-phase technology to achieve RNA synthesis at efficiencies rivaling those for DNA. This unit describes the synthesis of standard 5′-silyl-2′-ACE-protected phosphoramidites.

Keywords:

  • RNA synthesis;
  • silyl protection;
  • orthoester;
  • ribonucleoside;
  • phosphoramidite