title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides.
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.
authors.affiliation: Novartis Institutes for Biomedical Research (NIBR), Novartis Pharma AG, Basel, Basel-Stadt CH-4056, Switzerland and NIBR, Novartis Pharma AG, Cambridge, Massachusetts, MA 02139, USA.
authors.forename: Julien A
authors.initials: JA
authors.lastname: Boos
authors.forename: David W
authors.initials: DW
authors.lastname: Kirk
authors.forename: Mari-Luz
authors.initials: ML
authors.lastname: Piccolotto
authors.forename: Werner
authors.initials: W
authors.lastname: Zuercher
authors.forename: Sandro
authors.initials: S
authors.lastname: Gfeller
authors.forename: Philippe
authors.initials: P
authors.lastname: Neuner
authors.forename: Andre
authors.initials: A
authors.lastname: Dattler
authors.forename: William L
authors.initials: WL
authors.lastname: Wishart
authors.forename: Fabian
authors.initials: F
authors.lastname: Von Arx
authors.forename: Michael
authors.initials: M
authors.lastname: Beverly
authors.forename: Jesper
authors.initials: J
authors.lastname: Christensen
authors.forename: Karine
authors.initials: K
authors.lastname: Litherland
authors.forename: Esther
authors.initials: E
authors.lastname: van de Kerkhof
authors.forename: Pieter J
authors.initials: PJ
authors.lastname: Swart
authors.forename: Thomas
authors.initials: T
authors.lastname: Faller
authors.forename: Armin
authors.initials: A
authors.lastname: Beyerbach
authors.forename: David
authors.initials: D
authors.lastname: Morrissey
authors.forename: Juerg
authors.initials: J
authors.lastname: Hunziker
authors.forename: Iwan
authors.initials: I
authors.lastname: Beuvink
mesh_terms: D000818:Animals; D045325:HCT116 Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008297:Male; D051379:Mice; D009841:Oligonucleotides / Q000737:chemistry* / Q000493:pharmacokinetics / Q000627:therapeutic use; D009928:Organ Specificity; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics; D022661:RNA, Untranslated / Q000737:chemistry* / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014018:Tissue Distribution; D051598:Whole Body Imaging / Q000379:methods*
chemical_list: D009841:Oligonucleotides; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D022661:RNA, Untranslated