Multiplexed Intact-Tissue Transcriptional Analysis at Cellular Resolution

In recently developed approaches for high-resolution imaging within intact tissue, molecular characterization over large volumes has been largely restricted to labeling of proteins. But volumetric nucleic acid labeling may represent a far greater scientific and clinical opportunity, enabling detecti...

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Vydáno v:Cell Ročník 164; číslo 4; s. 792
Hlavní autoři: Sylwestrak, Emily Lauren, Rajasethupathy, Priyamvada, Wright, Matthew Arnot, Jaffe, Anna, Deisseroth, Karl
Médium: Journal Article
Jazyk:angličtina
Vydáno: United States 11.02.2016
Témata:
Adolescent
Animals
Brain Chemistry
Cyanates - chemistry
Ethyldimethylaminopropyl Carbodiimide - chemistry
Female
Humans
In Situ Hybridization - methods
Male
Maleimides - chemistry
Mice
Middle Aged
Nucleic Acid Amplification Techniques - methods
Oligonucleotides - chemistry
RNA - analysis
Succinimides - chemistry
Transcriptome
ISSN:1097-4172, 1097-4172
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Popis
Shrnutí:In recently developed approaches for high-resolution imaging within intact tissue, molecular characterization over large volumes has been largely restricted to labeling of proteins. But volumetric nucleic acid labeling may represent a far greater scientific and clinical opportunity, enabling detection of not only diverse coding RNA variants but also non-coding RNAs. Moreover, scaling immunohistochemical detection to large tissue volumes has limitations due to high cost, limited renewability/availability, and restricted multiplexing capability of antibody labels. With the goal of versatile, high-content, and scalable molecular phenotyping of intact tissues, we developed a method using carbodiimide-based chemistry to stably retain RNAs in clarified tissue, coupled with amplification tools for multiplexed detection. The resulting technology enables robust measurement of activity-dependent transcriptional signatures, cell-identity markers, and diverse non-coding RNAs in rodent and human tissue volumes. The growing set of validated probes is deposited in an online resource for nucleating related developments from across the scientific community.
Bibliografie:ObjectType-Article-1
SourceType-Scholarly Journals-1
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ISSN:1097-4172
1097-4172
DOI:10.1016/j.cell.2016.01.038