Design and Preparation of Aptamer-siRNA Chimeras (AsiCs) for Targeted Cancer Therapy

Methods Mol Biol. 2017:1632:175-186. doi: 10.1007/978-1-4939-7138-1_11.

Abstract

Conjugation of cell-internalizing DNA or RNA aptamers to tumor-suppressing siRNAs represents a novel promising approach for cancer therapy. Here we describe how to employ RNA aptamers that bind to cell surface receptors as carriers for cell-targeted siRNA (or miRNA) delivery. This protocol was optimized to improve the efficiency of the aptamer-siRNA/miRNA conjugates and facilitate its implementation in most molecular biology labs. The single working steps include (1) outlining the optimal sequences of the RNA strands bearing the aptamer and siRNA sequence, for which further (2) a dsDNA template is synthesized and then (3) transcribed into an RNA that will be (4) folded and annealed to a chemically synthesized siRNA complementary strand. Moreover, we reference recent examples and advances in the aptamer delivery field.

Keywords: Aptamers; In vitro transcription; Primer extension; RNAi; Targeted cancer therapy; Targeted delivery; miRNA; siRNA.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aptamers, Nucleotide*
  • Computational Biology / methods*
  • DNA
  • In Vitro Techniques
  • RNA Folding
  • RNA, Small Interfering / chemistry*
  • SELEX Aptamer Technique*
  • Templates, Genetic
  • Transcription, Genetic

Substances

  • Aptamers, Nucleotide
  • RNA, Small Interfering
  • DNA