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

Boos, Julien and Kirk, David William and Dattler, Andre and Piccolotto, Mari-Luz and Zuercher, Werner and Gfeller, Sandro and Neuner, Philippe and Christensen, Jesper and Wishart, William L. and Beverly, Michael and Von Arx, Fabian and Litherland, Karine and Van De Kerkhof, Esther and Swart, Pieter Jacob and Faller, Thomas and Beyerbach, Armin and Morrissey, David and Hunziker, Juerg and Beuvink, Iwan (2012) Whole-body scanning PCR; a sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides. Nucleic Acids Research, Nuclei. ISSN 0305-1048

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 RTqPCR 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.

Item Type: Article
Related URLs:
Keywords: siRNA,miRNA,mRNA,noncodingRNA,RTqPCR,Chemical-Ligation,Ligation,qPCR,whole body scanning PCR,RNA extraction
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Date Deposited: 13 Oct 2015 13:14
Last Modified: 13 Oct 2015 13:14
URI: https://oak.novartis.com/id/eprint/7984

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