Single-cell mRNA cytometry via sequence-specific nanoparticle clustering and trapping

Mahmoud Labib, Reza M. Mohamadi, Mahla Poudineh, Sharif U. Ahmed, Ivaylo Ivanov, Ching Lung Huang, Maral Moosavi, Edward H. Sargent, Shana O. Kelley*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

59 Scopus citations

Abstract

Cell-To-cell variation in gene expression creates a need for techniques that can characterize expression at the level of individual cells. This is particularly true for rare circulating tumour cells, in which subtyping and drug resistance are of intense interest. Here we describe a method for cell analysis-single-cell mRNA cytometry-That enables the isolation of rare cells from whole blood as a function of target mRNA sequences. This approach uses two classes of magnetic particles that are labelled to selectively hybridize with different regions of the target mRNA. Hybridization leads to the formation of large magnetic clusters that remain localized within the cells of interest, thereby enabling the cells to be magnetically separated. Targeting specific intracellular mRNAs enablescirculating tumour cells to be distinguished from normal haematopoietic cells. No polymerase chain reaction amplification is required to determine RNA expression levels and genotype at the single-cell level, and minimal cell manipulation is required. To demonstrate this approach we use single-cell mRNA cytometry to detect clinically important sequences in prostate cancer specimens.

Original languageEnglish (US)
Pages (from-to)489-495
Number of pages7
JournalNature chemistry
Volume10
Issue number5
DOIs
StatePublished - May 1 2018

ASJC Scopus subject areas

  • General Chemical Engineering
  • General Chemistry

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