High resolution spatial transcriptome analysis by photo-isolation chemistry
Mizuki HondaiD, Shinya OkiiD, Ryuichi KimuraiD, Akihito HaradaiD, Kazumitsu MaeharaiD, Kaori Y. TanakaiD, Chikara MenoiD, Yasuyuki OhkawaiD
Abstract
In multicellular organisms, individual cells are characterized by their gene expression profiles and the spatial interactions among cells enable the elaboration of complex functions. Expression profiling in spatially defined regions is crucial to elucidate cell interactions and functions. Here, we established a transcriptome profiling method coupled with photo-isolation chemistry (PIC) that allows the determination of expression profiles specifically from photo-irradiated regions of whole tissues. PIC uses photo-caged oligodeoxynucleotides for in situ reverse transcription. After photo-irradiation of limited areas, gene expression was detected from at least 10 cells in the tissue sections. PIC transcriptome analysis detected genes specifically expressed in small distinct areas of the mouse embryo. Thus, PIC enables transcriptome profiles to be determined from limited regions at a spatial resolution up to the diffraction limit.
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