Use of guanidinopropyl-modified siRNAs to silence gene expression

Methods Mol Biol. 2015:1218:217-49. doi: 10.1007/978-1-4939-1538-5_13.

Abstract

Silencing gene expression by harnessing the RNA interference (RNAi) pathway with short interfering RNAs (siRNAs) has useful analytical and potentially therapeutic application. To augment silencing efficacy of siRNAs, chemical modification has been employed to improve stability, target specificity, and delivery to target tissues. siRNAs incorporating guanidinopropyl (GP) moieties have demonstrated enhanced target gene silencing in cell culture and in vivo models of hepatitis B virus replication. Here we describe the synthesis of GP-modified siRNAs and use of 5' rapid amplification of cDNA ends (5' RACE) to verify an RNAi-mediated mechanism of action of these novel chemically modified siRNAs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Gene Expression
  • Gene Silencing*
  • Guanidines / chemistry
  • Hepatocytes / cytology
  • Hepatocytes / metabolism
  • Humans
  • Nucleic Acid Amplification Techniques
  • Oligonucleotides / chemical synthesis
  • Oligonucleotides / genetics*
  • Oligonucleotides / metabolism
  • Organophosphorus Compounds / chemistry
  • RNA, Small Interfering / chemical synthesis*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Solid-Phase Synthesis Techniques / methods*
  • Transfection

Substances

  • Guanidines
  • Oligonucleotides
  • Organophosphorus Compounds
  • RNA, Small Interfering
  • phosphoramidite