Fluorescent protein-based molecular beacons by zinc finger protein-guided assembly

Biotechnol Bioeng. 2015 Feb;112(2):236-41. doi: 10.1002/bit.25441. Epub 2014 Sep 29.

Abstract

Molecular beacons (MBs) are stem-loop structured oligonucleotides capable of sensitive and specific nucleic acid detection. However, large-scale usage of MBs for high throughput applications has been hindered by the many expensive and tedious chemical modifications. In this paper, we reported a new class of fluorescent protein-based molecular beacons (FP-MBs) based on zinc finger protein-guided assembly. The design consisted of a single oligonucleotide which forms a hairpin structure with an extending arm on both ends for the attachment of two different fluorescent proteins upon simple mixing. This new design allows for simplified MB assembly and modifications for a wide range of applications.

Keywords: detection; in vivo; mRNA.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cloning, Molecular
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Fluorescent Dyes / chemistry*
  • Fluorescent Dyes / metabolism
  • Luminescent Proteins / chemistry*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Molecular Probe Techniques
  • Molecular Probes / chemistry*
  • Molecular Probes / genetics
  • Molecular Probes / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Recombinant Fusion Proteins / chemistry*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Sensitivity and Specificity
  • Zinc Fingers / genetics*

Substances

  • Fluorescent Dyes
  • Luminescent Proteins
  • Molecular Probes
  • RNA, Messenger
  • Recombinant Fusion Proteins