Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides

Nucleic Acids Res. 2013 Aug;41(15):e145. doi: 10.1093/nar/gkt515. Epub 2013 Jun 13.

Abstract

Efficient tissue-specific delivery is a crucial factor in the successful development of therapeutic oligonucleotides. Screening for novel delivery methods with unique tissue-homing properties requires a rapid, sensitive, flexible and unbiased technique able to visualize the in vivo biodistribution of these oligonucleotides. Here, we present whole body scanning PCR, a platform that relies on the local extraction of tissues from a mouse whole body section followed by the conversion of target-specific qPCR signals into an image. This platform was designed to be compatible with a novel RT-qPCR assay for the detection of siRNAs and with an assay suitable for the detection of heavily chemically modified oligonucleotides, which we termed Chemical-Ligation qPCR (CL-qPCR). In addition to this, the platform can also be used to investigate the global expression of endogenous mRNAs and non-coding RNAs. Incorporation of other detection systems, such as aptamers, could even further expand the use of this technology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • HCT116 Cells
  • Humans
  • Image Processing, Computer-Assisted
  • Male
  • Mice
  • Oligonucleotides / chemistry*
  • Oligonucleotides / pharmacokinetics
  • Oligonucleotides / therapeutic use
  • Organ Specificity
  • Polymerase Chain Reaction / methods*
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics*
  • RNA, Small Interfering / chemistry
  • RNA, Small Interfering / genetics
  • RNA, Untranslated / chemistry*
  • RNA, Untranslated / genetics
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Tissue Distribution
  • Whole Body Imaging / methods*

Substances

  • Oligonucleotides
  • RNA, Messenger
  • RNA, Small Interfering
  • RNA, Untranslated