High-throughput intracellular biopsy of microRNAs for dissecting the temporal dynamics of cellular heterogeneity.
Journal Article (Journal Article)
The capability to analyze small RNAs responsible for post-transcriptional regulation of genes expression is essential for characterizing cellular phenotypes. Here, we describe an intracellular biopsy technique (inCell-Biopsy) for fast, multiplexed, and highly sensitive profiling of microRNAs (miRNAs). The technique uses an array of diamond nanoneedles that are functionalized with size-dependent RNA binding proteins, working as "fishing rods" to directly pull miRNAs out of cytoplasm while keeping the cells alive, thus enabling quasi-single-cell miRNA analysis. Each nanoneedle works as a reaction chamber for parallel in situ amplification, visualization, and quantification of miRNAs as low as femtomolar, which is sufficient to detect miRNAs of a single-copy intracellular abundance with specificity to single-nucleotide variation. Using inCell-Biopsy, we analyze the temporal miRNA transcriptome over the differentiation of embryonic stem cells (ESCs). The combinatorial miRNA expression patterns derived by inCell-Biopsy identify emerging cell subpopulations differentiated from ESCs and reveal the dynamic evolution of cellular heterogeneity.
Full Text
Duke Authors
Cited Authors
- Wang, Z; Qi, L; Yang, Y; Lu, M; Xie, K; Zhao, X; Cheung, EHC; Wang, Y; Jiang, X; Zhang, W; Huang, L; Wang, X; Shi, P
Published Date
- June 2020
Published In
Volume / Issue
- 6 / 24
Start / End Page
- eaba4971 -
PubMed ID
- 32577522
Pubmed Central ID
- PMC7286670
Electronic International Standard Serial Number (EISSN)
- 2375-2548
International Standard Serial Number (ISSN)
- 2375-2548
Digital Object Identifier (DOI)
- 10.1126/sciadv.aba4971
Language
- eng