title: Whole-Body Scanning PCR, a Tool for the Visualization of the In Vivo Biodistribution Pattern of Endogenous and Exogenous Oligonucleotides in Rodents.
abstract: Characterizing the in vivo biodistribution pattern and relative expression levels of oligonucleotide-based molecules such as mRNA, miRNA, siRNA, and anti-miRNAs in animal models, could be a helpful first-step in the successful development of therapeutic oligonucleotides. Here we describe a simple procedure called "Whole-Body Scanning PCR" (WBS-PCR), which combines the power of PCR with that of imaging. WBS-PCR relies on 384 well-defined extractions across a mouse whole-body section followed by a single dilution step which renders the lysates compatible with various qPCR-based assays. The in vivo biodistribution maps are generated by deconvoluting the qPCR data and converting it into a TissueView compatible image file which can be overlaid with an image of the whole-body section used for extractions. WBS-PCR is a flexible platform that can be adapted to other detection systems and thereby further expand the use of this technology.
authors.affiliation: Novartis Institutes for Biomedical Research (NIBR), Novartis Pharma AG, Fabrikstrasse 2, 4056, Basel, Switzerland.
authors.forename: Julien A
authors.initials: JA
authors.lastname: Boos
authors.affiliation: Novartis Institutes for Biomedical Research (NIBR), Novartis Pharma AG, Fabrikstrasse 2, 4056, Basel, Switzerland. iwan.beuvink@novartis.com.
authors.forename: Iwan
authors.initials: I
authors.lastname: Beuvink
mesh_terms: D000818:Animals; D016616:Cryoultramicrotomy / Q000379:methods; D015870:Gene Expression; D051379:Mice; D035683:MicroRNAs / Q000235:genetics; D023421:Models, Animal; D057054:Molecular Imaging / Q000379:methods*; D009841:Oligonucleotides / Q000008:administration & dosage / Q000235:genetics* / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012377:Rodentia; D014018:Tissue Distribution; D051598:Whole Body Imaging / Q000379:methods*
chemical_list: D035683:MicroRNAs; D009841:Oligonucleotides; D012333:RNA, Messenger; D012337:RNA, Ribosomal, 18S; D034741:RNA, Small Interfering
keywords: Imaging; In vivo biodistribution; Mouse; QWBA; RT-qPCR; Rat; Therapeutic oligonucleotide s; miRNA; qPCR; siRNA