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Nucleic Acids Res ; 41(15): e145, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23766292

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.


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